Christopher Shawhttps://scienceblogs.com/taxonomy/term/1449/feed
enAnother antivaccine paper bites the dusthttps://scienceblogs.com/insolence/2017/10/10/another-antivaccine-paper-bites-the-dust
<span>Another antivaccine paper bites the dust</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>I've written on quite a few occasions about a pair of scientists beloved by the antivaccine movement. I'm referring, of course, to <a href="http://respectfulinsolence.com/?s=Shaw+Tomljenovic">Christopher Shaw and Lucija Tomljenovic</a>. Whether it is their publishing dubious "evidence" that HPV vaccines cause <a href="http://respectfulinsolence.com/2016/03/01/one-more-time-theres-no-evidence-gardasil-causes-premature-ovarian-failure/">premature ovarian failure</a> or even <a href="http://respectfulinsolence.com/2012/10/31/and-now-death-by-gardasil-again-not-so-fast/">death</a> or <a href="http://respectfulinsolence.com/2011/12/08/and-global-warming-is-caused-by-the-decr/">demonizing aluminum as a vaccine adjuvant</a>, Shaw and Tomljenovic publish nothing but antivaccine pseudoscience that antivaxers love to cite whenever they dump some turd of a study on the medical literature.</p>
<!--more--><p>Just last month, they dumped their <a href="http://www.sciencedirect.com/science/article/pii/S0162013417300417#">latest turd of a study</a>, in which they basically tortured mice in the name of pseudoscience. Later, after I wrote my first analysis of the study in which I described how <a href="http://respectfulinsolence.com/2017/09/21/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-2017-aluminum-edition/">poorly designed and executed the experiments were</a>, I discovered that there's more than just bad science there. There's possible fraud, as circulating on PubPeer are reports of <a href="http://respectfulinsolence.com/2017/09/27/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-was-it-fraud-or-incompetence/">image manipulation that are quite convincing</a>. At the time this rather obvious image manipulation was being discussed, so, too, was the possibility of retraction. After all, if there's one thing that merits pretty much an automatic retraction in science, it's manipulation of images presented as data in a scientific paper.</p>
<p>Not surprisingly, then, yesterday I learned from Retraction Watch that Shaw and Tomljenovic's latest paper will be retracted as well. The editor of the <em>Journal of Inorganic Biochemistry</em> announced that the journal will be retracted:</p>
<blockquote><p>
The journal’s editor, John Dawson of the University of South Carolina, told Retraction Watch:</p>
<blockquote><p>
The paper by Shaw and co-workers is being retracted jointly by the authors and the editor.
</p></blockquote>
<p>He noted there will be a “statement accompanying the retraction of the paper.”</p>
<p>Shaw told us that his lab began investigating the issues raised on PubPeer “within a day” and reported its findings to both UBC and the journal soon after. He said:</p>
<blockquote><p>
Our own analysis showed some figures had been altered. We requested a retraction because we could not understand how that had happened. We felt the data had been compromised.
</p></blockquote>
<p>Shaw said that the problems mostly lie with data showing no change in gene or protein expression levels after aluminum injections — but also with some data showing changes in expression, which the paper attributed to the injections.
</p></blockquote>
<p>Next up, Shaw tries to pass the buck:</p>
<blockquote><p>
Shaw said that first author Dan Li, a former postdoc who performed the molecular biology and gene expression analysis for the study, has agreed to the retraction but not yet offered an official explanation about the data. Shaw told us:</p>
<blockquote><p>
She denied that anything had been manipulated, or that anything was amiss.
</p></blockquote>
<p>He added that when Li left the lab in 2015, she took the original data with her:</p>
<blockquote><p>
UBC policy is that original data never leave the lab. We’ve asked for them to be returned to us.
</p></blockquote>
<p>Shaw said he thinks the core data are “probably correct,” but said he plans to have the experiments re-done:</p>
<blockquote><p>
It is what it is. We’ve done everything we can on our end. We’re still having conversations with Li on where the data are and how we get them back. That’s as much as we can do at this point.
</p></blockquote>
</blockquote>
<p>I suppose that it's possible that Shaw was duped by a postdoctoral fellow in his laboratory. When you're the head of a lab and the principal investigator of a study, you tend to come to trust those working for you. You don't want to think that one of them might be committing scientific fraud by manipulating images. On the other hand, as PI, one has to be on guard for this very thing. The PI is basically the captain of the ship, and the buck stops at his desk, and whatever other cliche you want to invoke to say that he is in charge and responsible for the integrity of the data produced by his lab.</p>
<p>The kindest possible interpretation is that Christopher Shaw runs a loose ship, so loose that he didn't notice that many of the bands on the images of his DNA gels and the autoradiographs of his Western blots were duplicated, flipped, and otherwise manipulated. Certainly, letting the raw data and raw images out of the lab is not good lab practice, particularly in this day and age, when pretty much all images of gels and Western blots are recorded digitally. In my lab, for instance, there is a lab shared drive, and every single image generated is stored there, so that original images used to make figures can always be recovered. PRanoid PI that I am, I even periodically copy the whole shared drive to my own computer, which in turn is regularly backed up. Key figures are preserved on cloud drives.</p>
<p>The worst possible interpretation is that Shaw either knew about the image manipulation (or even ordered it) or that he put so much pressure on his postdoc to produce results that she felt that she had to falsify figures to produce what he wanted. Of course, I wonder about Shaw's practices. For instance, in my <a href="http://respectfulinsolence.com/2017/09/27/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-was-it-fraud-or-incompetence/">discussion of the image manipulation</a>, I noted that Shaw and Tomljenovic have at minimum engaged in self-plagiarism, recycling figures from a 2014 review article into which they dumped a little original data in their soon-to-be retracted paper. So, in terms of commonly accepted practices, we already know Shaw's rather...unconcerned. </p>
<p>Then, of course, there's Shaw and Tomljenovic's history. Back in 2015, they published a <a href="http://www.sciencedirect.com/science/article/pii/S0264410X16000165">paper purporting to show</a> that aluminum adjuvants in Gardasil caused behavioral abnormalities in young female mice that was <a href="https://www.skepticalraptor.com/skepticalraptorblog.php/retracted-hpv-vaccine-article-shaw-tomljenovic/">retracted</a> for this reason:</p>
<blockquote><p>
This article has been withdrawn at the request of the Editor-in-Chief due to serious concerns regarding the scientific soundness of the article. Review by the Editor-in-Chief and evaluation by outside experts, confirmed that the methodology is seriously flawed, and the claims that the article makes are unjustified. As an international peer-reviewed journal we believe it is our duty to withdraw the article from further circulation, and to notify the community of this issue.
</p></blockquote>
<p>Why is it that the University of British Columbia and its Department of Ophthalmology (which is the department where Shaw and Tomljenovic are based) put up with crap like this? Shaw must have tenure or pictures of the Dean having an affair. I can't think of any other reasons why he isn't long gone.</p>
<p>Ultimately the article was republished in a form that was nearly word-for-word identical version in an inferior journal. Such is the fate of antivaccine pseudoscience. It always comes back. That's why I liken it to zombies, Jason Voorhees, Michael Myers, Freddy Krueger, Chucky, or any other monsters that "die" at the end of one movie, only to inevitably reappear in a new sequel over and over and over again.</p>
<p>I have little doubt that Shaw and Tomljenovic's soon-to-be retracted paper will reappear somewhere else within a few months. It's what antivaccine scientists publishing dubious or even fraudulent research to promote the long discredited idea that vaccines cause autism do. Their pseudoscience never dies. It just keeps being recycled like the sequel to a 1980s slasher flick.</p>
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<span><a title="View user profile." href="https://scienceblogs.com/author/oracknows">oracknows</a></span>
<span>Mon, 10/09/2017 - 21:15</span>
Tue, 10 Oct 2017 01:15:24 +0000oracknows22639 at https://scienceblogs.comTorturing more mice in the name of antivaccine pseudoscience: PubPeer versus antivaxershttps://scienceblogs.com/insolence/2017/09/27/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-was-it-fraud-or-incompetence
<span>Torturing more mice in the name of antivaccine pseudoscience: PubPeer versus antivaxers</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>Last week, an antivaxer "challenged" me to look over <a href="http://www.sciencedirect.com/science/article/pii/S0162013417300417">a paper</a> purporting to show that aluminum adjuvants in vaccines cause inflammation of the brain and therefore contribute to autism, a paper that she would be "citing frequently." Being someone who lives by the motto, "be careful what you wish for," <a href="http://respectfulinsolence.com/2017/09/21/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-2017-aluminum-edition/">I looked it over in detail</a>. Not surprisingly, my conclusion was that the experiments were poorly done using obsolete and not very quantitative methodology and that the results do not support the conclusions made by the authors. I was not alone in this conclusion. Skeptical Raptor was, if anything, even <a href="https://www.skepticalraptor.com/skepticalraptorblog.php/aluminum-causes-autism-shaw-tomljenovic-vaccine/">harsher on the paper than I was</a>.</p>
<!--more--><p>The <a href="http://www.sciencedirect.com/science/article/pii/S0162013417300417">paper in question</a> came out of the lab of Christopher Shaw and Lucija Tomljenovic in the Department of Ophthalmology at the University of British Columbia. As I note every time I examine a paper by these two warriors for antivaccine pseudoscience, both have a long history of publishing antivaccine “research,” mainly falsely blaming the aluminum adjuvants in vaccines for autism and, well, just about any health problem children have and <a href="http://respectfulinsolence.com/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">blaming Gardasil for premature ovarian failure</a> and all manner of woes <a href="http://respectfulinsolence.com/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">up to and including death</a>. Shaw was even prominently featured in the rabidly antivaccine movie <a href="http://respectfulinsolence.com/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">The Greater Good</a>. Not surprisingly, they’ve <a href="http://respectfulinsolence.com/2016/02/17/no-gardasil-does-not-cause-behavioral-problems/">had a paper retracted</a>, as well.</p>
<p>Given the authors' history and a paper that I and others found completely consistent with that history of publishing bad science in the service of antivaccine views, you might reasonably ask: Why am I writing about it again? It turns out that I was indeed far too kind the first time around. You see, I didn't look at all the DNA gels and Western blot films closely enough. I confess that sometimes I don't, particularly when the images provided by the journal online are relatively low resolution. Fortunately, however, there are others with a much sharper eye for photos of DNA gels and films of Western blots than I am, and, if what these people are saying is correct, I rather suspect that Shaw and Tomljenovic might well be cruising for their second retracted paper. Before I explain why, it's necessary for me to briefly explain two things for nonscientists not familiar with the methodology used.</p>
<p>In last week's post, I complained that the authors had basically ground up mouse brains and used semiquantitative PCR to measure the level of messenger RNA for each immune cytokine examined. There's no need for me to go into how this method is only roughly quantitative or how there are much better methods available now. <a href="http://respectfulinsolence.com/2017/09/21/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-2017-aluminum-edition/">I did that last time</a>. What I do need to point out is that, after the PCR reaction is run, the PCR products (DNA fragments amplified by the PCR reaction) are separated by placing them in an agarose gel and running an electrical current through it. This gel electrophoresis works because DNA migrates towards the positive electrode and, once it solidifies, agarose forms a gel that separates the DNA fragments by size. The gel can then be stained with ethidium bromide, whose fluorescence allows visualization of the bands, which can be assessed for size and purity. Photos of the gel can be taken and subjected to densitometry to estimate how much DNA is in each band relative to the other bands.</p>
<p>To measure protein, Western blots work a little differently. Basically isolated cell extracts or protein mixtures are subjected to polyacrylamide gel electrophoresis (PAGE) with a denaturing agent (SDS). Again, like DNA, protein migrates towards the positive electrode, and the gel forms pores that impeded the process, allowing separation by size and charge. The proteins are then transferred to a membrane (the Western blot) and visualized by using primary antibodies to the desired protein, followed by a secondary antibody with some sort of label. In the old days, we often used radioactivity. These days, we mostly use chemiluminescence. Blots are then exposed to film or, more frequently today, to a phosphoimager plate, which provides a much larger linear range for detecting the chemiluminescence than old-fashioned film. Just like DNA gels, the bands can be quantified using densitometry. In both cases, it's very important not to "burn" (overexpose) the film, which pushes the band intensity out of the linear ranger) or to underexpose them (noise can cause problems). It's also important how the lines are drawn around the bands using the densitometry software and how the background is calculated. More modern software can do it fairly automatically, but there is almost always a need to tweak the outlines chosen, which is why I consider it important that whoever is doing the densitometry should be blinded to experimental group, as bias can be introduced in how the bands are traced.</p>
<p>So why did I go through all this? Hang on, I'll get to it. First, however, I like to point out to our antivaccine "friends" that peer review doesn't end when a paper is published. Moreover, social media and the web have made it easier than ever to see what other scientists think of published papers. In particular, there is a website called <a href="https://pubpeer.com">PubPeer</a>, which represents itself as an "online journal club." More importantly, for our purposes, PubPeer is a site where a lot of geeky scientists with sharp eyes for anomalies in published figures discuss papers and figures that seem, well, not entirely kosher. It turns out that some scientists with sharp eyes have been <a href="https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983">going over Shaw and Tomljenovic's paper</a>, and guess what? They've been finding stuff. In fact, they've been finding stuff that to me (and them) looks rather...suspicious.</p>
<p>One, for instance, took figure 1C of the paper and adjusted the background and contrast to accentuate differences in tones:</p>
<p><a href="https://scienceblogs.com/files/insolence/files/2017/09/image-1505914692638.jpg"><img src="http://scienceblogs.com/insolence/files/2017/09/image-1505914692638-450x226.jpg" alt="" width="450" height="226" class="aligncenter size-medium wp-image-11074" /></a></p>
<p>It was <a href="https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983#2">immediately noted:</a></p>
<blockquote><ol><li>A clear and deliberate removal of the Male 3 Control TNF result. This isn't an unacknowledged splice, as there is no background pattern from a gel contiguous with either band, left or right.</li>
<li>Removal of the left half of the Male 1 Control IFN-g. Dubious also about Male 3 Control IFN-g, as the contrast highlight shows boxing around the band.</li>
<li>What appears like an unacknowledged splice in ACHE blot, between AI Animal 2, Control Animal 3</li>
</ol><p>Comparing this representative blot to the densitometry accompanying it, they score from 5 independent experiments IFN-g fold change from control to AI, relative to actin, as on average 4.5, with an SEM ranging from ~2.7 to 6.5. This seems too good to be true.</p></blockquote>
<p>Look at the band. It's the second from last band. It looks as though the band has been digitally removed. There is an obvious square there. The edges are clear. Now, this could be a JPEG compression artifact. Indeed, one of the commenters is very insistent about reminding everyone that compression artifacts can look like a square and fool the unwary into thinking that some sort of Photoshopping had occurred.. However, I do agree with another of the PubPeer discussants this is enough of a problem that the journal should demand the original blot.</p>
<p>On this one, I'll give Shaw and Tomljenovic the benefit of the doubt. (Whether they deserve it or not, you can judge for yourself.) That might be a compression artifact. Other problems discovered in the gels are not so easily dismissed. For instance, there definitely appears to be the ol' duplicated and flipped gel bands trick going on in Figure 2A:</p>
<p><a href="https://scienceblogs.com/files/insolence/files/2017/09/RrLUlyk.jpg"><img src="http://scienceblogs.com/insolence/files/2017/09/RrLUlyk-450x296.jpg" alt="" width="450" height="296" class="aligncenter size-medium wp-image-11075" /></a></p>
<p>Spotting these takes a little bit of skill, but look for distinctive parts of bands and then look to see if they show up elsewhere. It's also necessary to realize that there could be multiple different exposures of the same band, such that the same band can appear more or less intense <em>and</em> mirror-imaged. You have to know what to look for, and I fear that some readers not familiar with looking at blots like these might not see the suspicious similarities, even when pointed out. Still, let's take a look. There are more examples, for instance, these two bands in Figure 4C:</p>
<p><a href="https://scienceblogs.com/files/insolence/files/2017/09/image-1506251227792.jpg"><img src="http://scienceblogs.com/insolence/files/2017/09/image-1506251227792-450x262.jpg" alt="" width="450" height="262" class="aligncenter size-medium wp-image-11076" /></a></p>
<p>And Figures 4B and 4D, where bubbles on the gels serve as markers:</p>
<p><a href="https://scienceblogs.com/files/insolence/files/2017/09/image-1506305957384.png"><img src="http://scienceblogs.com/insolence/files/2017/09/image-1506305957384-450x297.png" alt="" width="450" height="297" class="aligncenter size-medium wp-image-11077" /></a></p>
<p>You can look at the rest of the PubPeer images for yourself and decide if you agree that something fishy is going on here. I've seen enough that I think there is, as is <a href="https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983#16">pointed out near the end</a>:</p>
<blockquote><p>
Great to see such rapid progress being made: Band duplications firmly established for gels in Figs. 2 and 4. Perhaps we can add some RT-PCR from Fig. 1 too? In Fig. 2, seek out the band marked above that looks like a sailing boat with mast and forestay. Now look for it in Fig. 1A. And then perhaps check for any other duplications?</p></blockquote>
<p>Others note that Shaw and Tomljenovic have engaged in a bit of self-plagiarism, too. Figure 1 in the 2017 paper is identical (and I do mean identical, except that the bars in the older paper are blue) to a paper they published in 2014. Basically, they threw a little primary data into one of their crappy review articles trying to blame "environment" (i.e., vaccines) for autism, this one <a href="http://www.oapublishinglondon.com/article/1368">published in 2014 in OA Autism</a>. Don't take my word for it. Both articles are open-access, and you can judge for yourself.</p>
<p>Some <a href="https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983#20">comments from PubPeer</a>:</p>
<blockquote><p>
As far as I can see figure 1 is identical in the two papers? But in the 2014 paper hisograns are described as means +/- SEM from three independent experiments and in 2017 as means +/- SEM of five independent experiments? <a href="http://www.oapublishinglondon.com/article/1368">http://www.oapublishinglondon.com/article/1368</a></p></blockquote>
<p><a href="https://pubpeer.com/publications/4AEB7C8F30015079E2611157CF8983#21">And</a>:</p>
<blockquote><p>
Brazen self-plagiarism of the open access 2014 paper’s Fig. 1 is a key find by the human commentator. Especially since it is not in PubMed (though it is Ref. 166 here). This means that they have used certain elements of a single gel four times in three years: Nice work if you can get it.</p>
<p>Here is the direct link to 2014 Fig. 1</p>
<p><a href="http://www.oapublishinglondon.com/images/html_figures/1368_346.png">http://www.oapublishinglondon.com/images/html_figures/1368_346.png</a></p>
<p>The licence for the 2014 paper states “Creative Commons Attribution License (CC-BY)”. Unfortunately, the 2017 recycling of Fig. 1 is neither creative nor is it attributed.</p>
<p>What this means is that Elsevier were misled regarding the copyright situation and the originality of the work. So this finding surely gives the 2017 publisher a get out of jail card. If they choose to play it, they can now unilaterally withdraw this embarrassing Anti-vaxxer concoction on these grounds alone.</p>
<p>Don’t forget to archive the two papers for your records: They might disappear from the publishers’ web sites at some point.</p></blockquote>
<p>And that's <em>still</em> not all. Let's take a visit to our scaly friend, Skeptical Raptor, where he notes that <a href="https://themadvirologist.blogspot.com/2017/09/does-recent-paper-by-shaw-really-show.html">The Mad Virologist</a> and the <a href="https://scientistabe.wordpress.com/2017/09/25/neurosciencesjunk-sciences-autopsy-of-a-flawed-study-of-aluminum-and-brain-inflammation-li-et-al-j-inorg-biochem-2017/">Blood-Brain Barrier Scientist</a> jointly analyzed the paper and found:</p>
<blockquote><p>
But there are six other key points that limit what conclusions can be drawn from this paper:</p>
<ol><li>They selected genes based on old literature and ignored newer publications.</li>
<li>The method for PCR quantification is imprecise and cannot be used as an absolute quantification of expression of the selected genes.</li>
<li>They used inappropriate statistical tests that are more prone to giving significant results which is possibly why they were selected.</li>
<li>Their dosing regime for the mice makes assumptions on the development of mice that are not correct.</li>
<li>They gave the mice far more aluminum sooner than the vaccine schedule exposes children to.</li>
<li>There are irregularities in both the semi-quantitative RT-PCR and Western blot data that strongly suggests that these images were fabricated. This is probably the most damning thing about the paper. If the data were manipulated and images fabricated, then the paper needs to be retracted and UBC needs to do an investigation into research misconduct by the Shaw lab.</li>
</ol><p>Taken together, we cannot trust Shaw’s work here and if we were the people funding this work, we’d be incredibly ticked off because they just threw away money that could have done some good but was instead wasted frivolously. Maybe there’s a benign explanation for the irregularities that we’ve observed, but until these concerns are addressed this paper cannot be trusted.</p></blockquote>
<p>I note that they go into even more detail about the problems with the images that have led me (and others) to be suspicious of image manipulation, concluding:</p>
<blockquote><p>
These are some serious concerns that raise the credibility of this study and can only be addressed by providing a full-resolution (300 dpi) of the original blots (X-ray films or the original picture file generated by the gel acquisition camera).</p>
<p>There has been a lot of chatter on PubPeer discussing this paper and many duplicated bands and other irregularities have been identified by the users there. If anyone is unsure of how accurate the results are, we strongly suggest looking at what has been identified on PubPeer as it suggests that the results are not entirely accurate and until the original gels and Western blots have been provided, it looks like the results were manufactured in Photoshop.</p></blockquote>
<p>I agree. Oh, and I agree with their criticism of the use of statistics. I even brought up their failure to control for multiple comparisons, but Shaw and Tomljenovic also used a test that is appropriate for a normal distribution when their data obviously did not follow a normal distribution.</p>
<p>So, my dear readers, it turns out that Orac, as Insolent as he can be when slapping down bad science by antivaxers, was not nearly Insolent enough in this case. Mea culpa. I should have known better, given Shaw and Tomljenovic's history. Not only do we have poorly done and analyzed experiments, but we also have self-plagiarism and, quite possibly, scientific fraud. Only releasing the full resolution original images from the original experiments (which are now probably four years old) can put these questions to rest.</p>
<p>Science matters. I hate to see it abused like this, particularly when experimental animals are killed in the service of such awful science.</p>
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<span>Wed, 09/27/2017 - 01:00</span>
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Wed, 27 Sep 2017 05:00:40 +0000oracknows22631 at https://scienceblogs.comTorturing more mice in the name of antivaccine pseudoscience, 2017 aluminum editionhttps://scienceblogs.com/insolence/2017/09/21/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-2017-aluminum-edition
<span>Torturing more mice in the name of antivaccine pseudoscience, 2017 aluminum edition</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><div class="featured-image"><img alt="" class="attachment-post-thumbnail size-post-thumbnail wp-post-image" data-entity-type="" data-entity-uuid="" height="344" sizes="(max-width: 590px) 100vw, 590px" src="http://scienceblogs.com/insolence/files/2016/11/Mousetorture-590x344.jpg" width="590" /><div class="caption" style="width:590px;"> </div>
<div class="caption" style="width:590px;">"Why, oh, why do I have to die in the cause of such crappy science?"</div>
</div>
<p>For antivaxers, aluminum is the new mercury.</p>
<p>Let me explain, for the benefit of those not familiar with the antivaccine movement. For antivaxers, it is, first and foremost, always about the vaccines. Always. Whatever the chronic health issue in children, vaccines must have done it. Autism? It’s the vaccines. Sudden infant death syndrome? Vaccines, of course. Autoimmune diseases? Obviously it must be the vaccines causing it. Obesity, diabetes, ADHD? Come on, you know the answer!</p>
<p>Because antivaxers will never let go of their obsession with vaccines as The One True Cause Of All Childhood Health Problems, the explanation for how vaccines supposedly cause all this harm are ever morphing in response to disconfirming evidence. Here’s an example. Back in the late 1990s and early 2000s, antivaxers in the US (as opposed to in the UK, where the MMR vaccine was the bogeyman) focused on mercury in vaccines as the cause of autism. That’s because many childhood vaccines contained thimerosal, a preservative that contains mercury. In an overly cautious bit of worshiping at the altar of the precautionary principle, in 1999 the CDC recommended removing the thimerosal from childhood vaccines, and as a result it was removed from most vaccines by the end of 2001. (Some flu vaccines continued to contain thimerosal for years after that, but no other childhood vaccine did, and these days it’s uncommon for thimerosal-containing vaccines of any kind.)</p>
<p>More importantly, the removal of thimerosal from childhood vaccines provided a natural experiment to test the hypothesis that mercury causes or predisposes to autism. After all, if mercury in vaccines caused autism, the near-complete removal of that mercury from childhood vaccines in a short period of time should have resulted in a decline in autism prevalence beginning a few years after the removal. Guess what happened? Autism prevalence didn’t decline. It continued to rise. To scientists, this observation was a highly convincing falsification of the hypothesis through a convenient natural experiment, although those who belong to the strain of antivaccine movement sometimes referred to as the mercury militia still flog mercury as a cause of autism even now. Robert F. Kennedy, Jr. is perhaps the most famous mercury militia member, although of late he’s been sounding more and more like a run-of-the-mill antivaxer.</p>
<p>Which brings us to aluminum.</p>
<p>With mercury in vaccines pretty definitively eliminated as The One True Cause Of Autism, antivaxers started looking for other ingredients to blame for autism because, as I said before, it’s first, foremost, and always all about the vaccines. So naturally they shifted their attention to the aluminum adjuvants in many vaccines. Adjuvants are compounds added to vaccine in order to boost the immune response to the antigen used, and aluminum salts <a href="https://www.facebook.com/RtAVM/posts/10152027115158831">have been used</a> as <a href="http://vk.ovg.ox.ac.uk/vaccine-ingredients#aluminium">effective adjuvants</a> for <a href="https://www.cdc.gov/vaccinesafety/concerns/adjuvants.html">many years now</a> and <a href="http://www.chop.edu/centers-programs/vaccine-education-center/vaccine-ingredients/aluminum">have an excellent safety record</a>. None of that has stopped antivaxers from trying to make aluminum the new mercury by blaming aluminum-containing vaccines for autism. I was reminded by this earlier this week when my e-mail was flooded with messages about new study being <a href="https://medium.com/@jbhandley/new-canadian-study-autism-aluminum-adjuvant-link-corroborated-330e947f5f62" rel="nofollow">flogged by antivaxers</a> in spectacularly ignorant ways, including three—yes, three—identical messages from a certain antivaxer with a severe case of Dunning-Kruger and delusions of grandeur basically challenging me to review this study and assuring me that antivaxers would be citing it for a long time. Well, whenever I receive messages like that, particularly annoying repetition, my answer is: Be very careful what you wish for.</p>
<p>Also: Challenge accepted.</p>
<p>Which brings us to the study itself. It’s by antivaccine “researchers” whose <a href="http://respectfulinsolence.com/2016/11/18/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience/">previous studies</a> and <a href="http://respectfulinsolence.com/2011/12/08/and-global-warming-is-caused-by-the-decr/">review articles</a> I’ve <a href="http://respectfulinsolence.com/2016/02/17/no-gardasil-does-not-cause-behavioral-problems/">discussed before</a>. Yes, I’m referring to Christopher Shaw and Lucija Tomljenovic in the Department of Ophthalmology at the University of British Columbia. Both have a long history of publishing antivaccine “research,” mainly falsely blaming the aluminum adjuvants in vaccines for autism and, well, just about any health problem children have and <a href="http://respectfulinsolence.com/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">blaming Gardasil for premature ovarian failure</a> and all manner of woes <a href="http://respectfulinsolence.com/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">up to and including death</a>. Shaw was even prominently featured in the rabidly antivaccine movie <a href="http://respectfulinsolence.com/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">The Greater Good</a>. Not surprisingly, they’ve <a href="http://respectfulinsolence.com/2016/02/17/no-gardasil-does-not-cause-behavioral-problems/">had a paper retracted</a>, as well..</p>
<p>This time around, they’ve gone back to their old stomping grounds, the <em>Journal of Inorganic Biochemistry</em>, and, along with two other co-authors, published <a href="http://www.sciencedirect.com/science/article/pii/S0162013417300417#">Subcutaneous injections of aluminum at vaccine adjuvant levels activate innate immune genes in mouse brain that are homologous with biomarkers of autism</a>. It’s where they published review article in 2011 <a href="http://respectfulinsolence.com/2011/12/08/and-global-warming-is-caused-by-the-decr/">full of antivaccine misinformation and distortions</a>. So, given Shaw and Tomljenovic’s history, it is not unreasonable to be suspicious of this study as well. But, hey, you never know. Maybe it’s a good study that sheds light on an important aspect of the pathogenesis of autism…Ah, who’m I kidding? It’s nothing of the sort. It’s yet another study designed to imply that aluminum adjuvants cause autism.</p>
<p>Before we look at the study itself, specifically the experiments included in it, let’s consider the hypothesis being tested, because experiments in any study should be directed at falsifying the hypothesis. Unfortunately, there is no clear statement of hypothesis where it belongs, namely in the introduction. Instead, what we get is this:</p>
<blockquote><p>Given that infants worldwide are regularly exposed to Al adjuvants through routine pediatric vaccinations, it seemed warranted to reassess the neurotoxicity of Al in order to determine whether Al may be considered as one of the potential environmental triggers involved in ASD.</p>
<p>In order to unveil the possible causal relationship between behavioral abnormalities associated with autism and Al exposure, we initially injected the Al adjuvant in multiple doses (mimicking the routine pediatric vaccine schedule) to neonatal CD-1 mice of both sexes.</p>
</blockquote>
<p>This is basically a fishing expedition in which the only real hypothesis is that “aluminum in vaccines is bad and causes bad immune system things to happen in the brain.” “Fishing expeditions” in science are studies in which the hypothesis is not clear and the investigators are looking for some sort of effect that they suspect they will find. In fairness, fishing expeditions are not a bad thing in and of themselves—indeed, they are often a necessary first step in many areas of research—but they are hypothesis-generating, not hypothesis confirming. After all, there isn’t a clear hypothesis to test; otherwise it wouldn’t be a fishing expedition. The point is that this study does not confirm or refute any hypothesis, much less provide any sort of slam-dunk evidence that aluminum adjuvants cause autism.</p>
<p>Moving along, I note that this is a mouse experiment, and somehow antivaxers are selling this as compelling evidence that vaccines cause autism through their aluminum adjuvants causing an inflammatory reaction in the brain. Now, seriously. Mouse models can be useful for a lot of things, but, viewed critcally, for the most part autism is not really one of them. After all, autism is a human neurodevelopmental disorder diagnosed entirely by behavioral changes, and correlating mouse behavior with human behavior is very problematic. Indeed, correlating the behavior of any animal, even a primate, with human behavior is fraught with problems. Basically, there is no well-accepted single animal model of autism, and autism research has been littered with mouse models of autism that were found to be very much wanting. (<a href="http://photoninthedarkness.blogspot.com/2005/07/dr-hornigs-autistic-mice_29.html">“Rain mouse,” anyone?</a>) Basically, despite the <a href="https://spectrumnews.org/opinion/viewpoint/promises-and-limitations-of-mouse-models-of-autism/">existence of many mouse strains</a> touted to be relevant to autism, almost none of them are truly relevant because:</p>
<blockquote><p>A good animal model satisfies three fundamental criteria. The first, called face validity, requires sufficient similarities between the phenotype of the mice and symptoms of the human disorder. The second, called construct validity, is achieved if the biological cause of the human disease is replicated in the mouse — for example, when an autism-associated gene is mutated in mice. Finally, a mouse model has predictive validity if treatments improve both the human symptoms of the disorder and the mouse phenotype.</p>
<p>Diagnosis of autism is purely behavioral and requires clearly defined symptoms in each of three core categories: abnormal social interactions, impaired communication and repetitive behavior. One of the challenges in studying mouse models is determining which behaviors from the mouse repertoire could be considered analogous to these symptoms.</p>
</blockquote>
<p>And:</p>
<blockquote><p>So far, very few of these mouse models display behavioral phenotypes relevant to all three core domains of autism. What’s more, in some cases, physical problems such as poor general health following seizures, or low exploratory activity, produce false positives that prevent the interpretation of more complex, autism-relevant phenotypes.</p>
</blockquote>
<p>Pay particular attention to the part about construct validity. The assumption behind this study is that immune changes in the brain of mice will be relevant to immune activation in the brains of autistic humans. That is an assumption that hasn’t yet been confirmed with sufficient rigor to view this study’s results as any sort of compelling evidence that aluminum adjuvants cause autism. Yes, the authors include this important-looking diagram describing how they think immune system activation causes autism (click to embiggen):</p>
<p><a href="http://scienceblogs.com/insolence/files/2017/09/InflammationAutism.jpg"><img alt="" class="aligncenter size-medium wp-image-11064" data-entity-type="" data-entity-uuid="" height="184" sizes="(max-width: 450px) 100vw, 450px" src="http://scienceblogs.com/files/insolence/files/2017/09/InflammationAutism-450x184.jpg" /></a></p>
<p>In the end, though, as impressive as it is, the relevance of this chart to autism is questionable at best, as is the relevance of this study. So let’s look at the mouse strain chosen by the investigators, <a href="http://www.criver.com/files/pdfs/rms/cd1/rm_rm_d_cd1_mouse.aspx">CD-1 mice</a>. Basically, there’s nothing particularly “autistic” (even in terms of existing mouse models purported to be relevant to autism) about these mice, which are described in most catalogues of companies selling them as “general purpose.” Basically, the authors used them because they had used them before in previous studies in which they reported that aluminum injections caused <a href="http://www.sciencedirect.com/science/article/pii/S0162013409001809">motor neuron degeneration</a> (nope, no autism) and another crappy paper in the same journal from 2013 purporting to <a href="http://www.sciencedirect.com/science/article/pii/S0162013413001773">link aluminum with adverse neurological outcomes</a>. That’s it.</p>
<p>As for the experiment itself, neonatal mice were divided into two groups, a control group that received saline injections and the experimental group received injections of aluminum hydroxide in doses timed such that they that purportedly mimicked the pediatric vaccine schedule. Looking over the schedule used, I can’t help but note that there’s a huge difference between human infant development and mouse development. Basically, the mice received aluminum doses claimed to be the same as what human babies get by weight six times in the first 17 days of life. By comparison, in human babies these doses are <a href="https://www.vaccines.gov/who_and_when/infants_to_teens/index.html">separated by months</a>. In addition, in human babies, vaccines are injected intramuscularly (in a muscle). In this study, the mice were injected subcutaneously (under the skin). This difference immediately calls into question applicability and construct validity. The authors stated that they did it because they wanted to follow previously utilized protocols in their laboratory. In some cases, that can be a reasonable rationale for an experimental choice, but in this case the original choice was questionable in the first place. Blindly sticking with the same bad choice is just dumb.</p>
<p>So what were the endpoints examined in the mice injected with aluminum hydroxide compared to saline controls? After 16 weeks, the mice were euthanized and their brains harvested to measure gene expression and the levels of the proteins of interest. Five males and five females from each group were “randomly paired” for “gene expression profiling.” Now, when I think of gene expression profiling, I usually think of either cDNA microarray experiments, in which the levels of thousands of genes are measured at the same time, or next generation sequencing, in which the level of every RNA transcript in the cell can be measured simultaneously. That doesn’t appear to be what the authors did. Instead, they used a technique known as PCR to measure the messenger RNA levels of a series of cytokines. Basically, they examined the amount of RNA coding for various immune proteins in the brain chosen by the authors as relevant to inflammation. The authors also did Western blots for many of those proteins, which is a test in which proteins are separated on a gel, blotted to a filter, and then probed with specific antibodies, resulting in bands that can be measured by a number of techniques, including autoradiography or chemiluminescence, both of which can be recorded on film on which the relevant bands can be visualized. Basically, what the authors did wasn’t really gene expression profiling. It was measuring a bunch of genes and proteins and hoping to find a difference.</p>
<p>There’s an even weirder thing. The authors didn’t use quantitative real time reverse transcriptase PCR, which has been the state-of-the-art for measuring RNA message levels for quite some time. Rather, they used a very old, very clunky form of PCR that can only produce—at best—semiquantitative results. (That’s why we used to call it semiquantitative PCR.) Quite frankly, in this day and age, there is absolutely zero excuse for choosing this method for quantifying gene transcripts. If I were a reviewer for this article, I would have recommended not publishing it based on this deficiency alone. Real time PCR machines, once very expensive and uncommon, are widely available. (Hell, I managed to afford very simple one in my lab nearly 15 years ago.) Any basic or translational science department worth its salt has at least one available to its researchers.</p>
<p>The reason that this semiquantitative technique is considered inadequate is that the amount of PCR product grows exponentially, roughly doubling with every cycle of PCR, asymptotically approaching a maximum as the primers are used up.<br />
It usually takes around 30-35 cycles before everything saturates and the differences observed in the intensity of the DNA bands when they are separated on a gel become indistinguishable. That’s why PCR was traditionally and originally primarily considered a “yes/no” test. Either the RNA being measured was there and produced a PCR band, or it didn’t. In this case, the authors used 30 cycles, which is more than enough to result in saturation. (Usually semiquantitative PCR stops around 20-25 cycles or even less.) And I didn’t even (yet) mention how the authors didn’t use DNAse to eliminate the small amounts of DNA that contaminate nearly all RNA isolations. Basically, the primers used for PCR pick up DNA as well as any any RNA, and DNA for the genes of interest will be guaranteed to contaminate the specimens without DNAse treatment. Yes, you molecular biologists out there, I know that’s simplistic, but my audience doesn’t consist of molecular biologists.</p>
<p>Now, take a look at Figures 1A and 1B as well as Figures 2A and 2B. (<a href="http://www.sciencedirect.com/science/article/pii/S0162013417300417#t0015">You can do it if you want</a>. The article is open access.) Look at the raw bands in the A panels of the figures. Do you see much difference, except for IFNG (interferon gamma) in Figure 1A? I don’t. What I see are bands of roughly the same intensity, even the ones that are claimed to vary by three-fold. In other words, I basically am very skeptical that the investigators saw much of difference in gene expression between controls and the aluminum-treated mice. In fairness, for the most part, the protein levels as measured by Western blot did correlate with what was found on PCR, but there’s another odd thing. The investigators didn’t do Western blots for all the same proteins whose gene expression they measured by PCR. Of course, they present primers for 27 genes, but only show blots for 18 (17 inflammatory genes plus beta actin, which was used as a standard to normalize the values for the other 17 genes).</p>
<p>I also question the statistical tests chosen by the authors. Basically, they examined each gene separately and used Student’s t-test to assess statistical significance. However, in reality they did many comparisons, at least 17, and there’s no evidence that the authors controlled for multiple comparisons. If one chooses statistical significance to occur at p < 0.05 and compares 20 samples, by random chance alone at least one will be different. Add to that the fact that there is no mention of whether the people performing the assays were blinded to experimental group, and there's a big problem. Basic science researchers often think that blinding isn't necessary in their work, but there is a potential for unconscious bias that they all too often don't appreciate. For example, the authors used Image J, free image processing software developed by the NIH. I've used Image J before. It's a commonly used app used to quantify the density of bands on gels, even though it's old software and hasn't been updated in years. Basically, it involves manually drawing outlines of the bands, setting the background, and then letting the software calculate the density of the bands. The potential for bias shows up in how you draw the lines around the bands and set the backgrounds. As oblivious as they seem to be to this basic fact, basic scientists are just as prone to unconscious bias as the rest of us, and, absent blinding, in a study like this there is definitely the potential for unconscious bias to affect the results. In fairness, few basic science researchers bother to blind whoever is quantifying Western blots or ethidium bromide-stained DNA gels of PCR products, but that's just a systemic problem in biomedical research that I not infrequently invoke when I review papers. Shaw and Tomljenovic are merely making the same mistake that at least 90% of basic scientists make.</p>
<p>But let’s step back and take the authors’ results at face value for a moment. Let’s assume that what is reported is a real effect. In the rest of the paper, the authors present evidence of changes in gene expression that suggest the activation of a molecular signaling pathway controlled by a molecule called NF-κB and that male mice were more susceptible to this effect than females. (Just like autism!) Funny, but I know NF-κB. I’ve <a href="https://www.ncbi.nlm.nih.gov/pubmed/20421348">published on NF-κB</a>. I had an NIH R01 grant to study how my favorite protein affected NF-κB. True, I ended up abandoning that line of research because I hit some dead ends. True, I’m not as familiar with NF-κB as I used to be. But I do know enough to know that NF-κB is easy to activate and very nonspecific. I used to joke that just looking at my cells funny would activate NF-κB signaling. Also, NF-κB activation is indeed associated with inflammation, but so what? What we have is an artificial model in which the mice are dosed much more frequently with aluminum than human infants. Does this have any relevance to the human brain or to human autism? who knows? Probably not. No, almost certainly not.</p>
<p>Also, the mouse immune system is <a href="https://www.ncbi.nlm.nih.gov/pubmed/14978070">different</a> from the <a href="https://www.ncbi.nlm.nih.gov/pubmed/25404048">human</a> <a href="http://www.pnas.org/content/110/8/2946.full.pdf">immune system</a>. None of this stops the authors from concluding:</p>
<blockquote><p>Based on the data we have obtained to date, we propose a tentative working hypothesis of a molecular cascade that may serve to explain a causal link between Al and the innate immune response in the brain. In this proposed scheme, Al may be carried by the macrophages via a Trojan horse mechanism similar to that described for the human immunodeficiency virus (HIV) and hepatitis C viruses, travelling across the blood-brain-barrier to invade the CNS. Once inside the CNS, Al activates various proinflammatory factors and inhibits NF-κB inhibitors, the latter leading to activation of the NF-κB signaling pathway and the release of additional immune factors. Alternatively, the activation of the brain’s immune system by Al may also occur without Al traversing the blood-brain barrier, via neuroimmuno-endocrine signaling. Either way, it appears evident that the innate immune response in the brain can be activated as a result of peripheral immune stimuli. The ultimate consequence of innate immune over-stimulation in the CNS is the disruption of normal neurodevelopmental pathways resulting in autistic behavior.</p>
</blockquote>
<p>That’s what we call in the business conclusions not supported by the findings in a study. On a more “meta” level, it’s not even clear whether the markers of inflammation observed in autistic brains are causative or an epiphenomenon. As Skeptical Raptor noted. It could be that the inflammation reported is caused by whatever the primary changes in the brain that result in autism. Cause and effect are nowhere near clear. One can’t help but note that many of the infections vaccinated against cause way more activation of the immune system and cytokines than vaccination.</p>
<p>So what are we left with?</p>
<p>Basically, what we have is yet another mouse study of autism. The study purports to show that aluminum adjuvants cause some sort of “neuroinflammation,” which, it is assumed, equals autism. By even the most charitable interpretation, the best that can be said for this study is that it might show increased levels of proteins associated with inflammation in the brains of mice who had been injected with aluminum adjuvant way more frequently than human babies ever would be. Whether this has anything to do with autism is highly questionable. At best, what we have here are researchers with little or no expertise in very basic molecular biology techniques using old methodology that isn’t very accurate overinterpreting the differences in gene and protein levels that they found. At worst, what we have are antivaccine “researchers” who are not out for scientific accuracy but who actually want to promote the idea that vaccines cause autism. (I know, I know, it’s hard not to ask: Why not both?) If this were a first offense, I’d give Shaw and Tomljenovic the benefit of the doubt, but this is far from their first offense. Basically, this study adds little or nothing to our understanding of autism or even the potential effects of aluminum adjuvants. It was, as so many studies before, the torture of mice in the name of antivax pseudoscience. <a href="https://www.skepticalraptor.com/skepticalraptorblog.php/aluminum-causes-autism-shaw-tomljenovic-vaccine/">The mice used in this study died in vain</a> in a study supported by the <a href="http://respectfulinsolence.com/2017/05/24/is-tipper-gore-appearing-at-a-fundraiser-hosted-by-antivaxers/">profoundly antivaccine Dwoskin Foundation</a>.</p>
<p>Also, I’ll tell my antivax admirer the same thing I once told J.B. Handley when he taunted me to examine a study that he viewed as “slam dunk” evidence for a vaccine-autism link: <a href="http://respectfulinsolence.com/2008/05/17/some-monkey-business-in-autism-research/">You don’t tug on Superman’s cape</a>. And, no, <a href="https://www.azlyrics.com/lyrics/jimcroce/youdontmessaroundwithjim.html">your name isn’t Slim</a>. You’re not an exception.</p>
<p><strong>ADDENDUM 9/27/2017</strong>: Apparently I wasn’t…Insolent…enough with this paper. On PubPeer there is a big discussion about whether the images in this paper were manipulated and whether the authors self-plagiarized Figure 1 from another paper. <a href="http://respectfulinsolence.com/2017/09/27/torturing-more-mice-in-the-name-of-antivaccine-pseudoscience-was-it-fraud-or-incompetence/">It looks bad</a>.</p>
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Thu, 21 Sep 2017 05:00:11 +0000oracknows22627 at https://scienceblogs.comMore Gardasil fear mongering: A "critical review" of HPV vaccination that lacks critical thinkinghttps://scienceblogs.com/insolence/2017/08/16/more-gardasil-fear-mongering-a-critical-review-of-hpv-vaccination-that-lacks-critical-thinking
<span>More Gardasil fear mongering: A "critical review" of HPV vaccination that lacks critical thinking</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>Here we go again.</p>
<p>Antivaxers don't like vaccines. This, we know. They blame them for everything from autism to autoimmune diseases to diabetes to sudden infant death syndrome. They even sometimes claim that shaken baby syndrome is a "misdiagnosis" for vaccine injury. However, there are two vaccines that stand out above all as the objects of antivaccine scorn. the first, of course, is the MMR vaccine. That's on Andrew Wakefield., of course, who almost singlehandedly popularized the fear that the MMR vaccine causes autism. The second most hated vaccine (by antivaxers) is Gardasil or Cervarix, both vaccines against the human papilloma virus (HPV), the virus that causes genital wart and certain subtypes of which greatly predispose to cancer. The reason for the extreme negative reaction to HPV vaccines is not because they are thought to cause autism, given that they are not administered until girls are approaching pubert. Rather, it is because they are falsely considered to be unnecessary. HPV is sexually transmitted disease, which makes the cervical cancer HPV vaccines prevent a largely sexually transmitted disease. Many religiously inclined antivaxers falsely think that HPV vaccination will encourage promiscuity, even though the evidence is pretty <a href="https://sciencebasedmedicine.org/no-the-hpv-vaccine-does-not-cause-promiscuity/">strong that it doesn't</a>. Nor does it cause <a href="http://scienceblogs.com/insolence/2016/03/01/one-more-time-theres-no-evidence-gardasil-causes-premature-ovarian-failure/">premature ovarian failure</a> or <a href="http://scienceblogs.com/insolence/2012/10/31/and-now-death-by-gardasil-again-not-so-fast/">sudden death</a>, as antivaxers frequently claim. Nor is it <a href="http://scienceblogs.com/insolence/2012/03/14/beware-vaccines-will-sap-and-impurity-yo/">contaminating your precious bodily fluids with HPV DNA</a>.</p>
<!--more--><h2>In which Orac learns of a "horrifying" review article about HPV vaccination</h2>
<p>So it was with little surprise at all that yet another allegedly systematic review article attempting to demonstrate that HPV vaccines are not safe. (<a href="https://www.skepticalraptor.com/skepticalraptorblog.php/one-stop-shop-science-myth-debunking-gardasil/">They are</a>.) I've been down this road before, but I do tend to take a look when new examples of these sorts of studies roll around because I never know what I am going to find and also know that these sorts of studies tend to get shared in a rabidly viral fashion on social media by antivaxers. If I can get my take on them out before that happens, then at least there's something out there to combat the misinformation. I didn't quite get there in that our old buddy "Dr. Jay" Gordon posted a link to Twitter:</p>
<blockquote class="twitter-tweet" data-lang="en"><p lang="en" dir="ltr" xml:lang="en" xml:lang="en">Meta-analysis reveals ‘worrisome’ results from HPV vaccine trials </p>
<p>This is a pharmaceutical industry journal <a href="https://t.co/FEjEyek1SN">https://t.co/FEjEyek1SN</a></p>
<p>— Jay Gordon, MD, FAAP (@JayGordonMDFAAP) <a href="https://twitter.com/JayGordonMDFAAP/status/897489148684025857">August 15, 2017</a></p></blockquote>
<script async="" src="//platform.twitter.com/widgets.js" charset="utf-8"></script><p>
On the other hand, Dr. Jay did tip me off to the study. So I suppose I should thank him for that. In any case, let's first look at how the article is being reported. The link above is to <a href="http://www.epmmagazine.com/news/meta-analysis-reveals-‘worrisome’-results-from-hpv-vaccine-t/">European Pharmaceutical Manufacturer</a> (epm) a magazine that really should know better:</p>
<blockquote><p>
The study, performed by researchers from Mexico’s National Institute of Cardiology and published online in Clinical Rheumatology, identified randomised trials of HPV vaccines in PubMed and reviewed post-marketing case series highlighting serious adverse events of the vaccines.</p>
<p>It was found that in the majority of the randomised studies identified the control groups were not administered with an inert placebo but rather an aluminium placebo. According to a report published by <a href="http://www.collective-evolution.com/2017/08/12/new-study-vaccine-manufacturers-fda-regulators-caught-hiding-risks-of-hpv-vaccines/">Collective Evolution</a> using an aluminium based placebos would offer up much different comparative results than when administering a pharmacologically inert placebo.</p>
<p>When comparing the rates of serious adverse events, the researchers found that in two of the largest randomised trials, there were more occurrences in the HPV treated groups than in the aluminium placebo groups. Additionally, comparing girls vaccinated with the 4-valent HPV vaccine and those receiving the 9-valent dose, the latter group had more serious systemic adverse events. The researchers also revealed a ‘worrisome’ quotient of the number needed to vaccinate/number needed to harm in the nine-valent HPV vaccine.
</p></blockquote>
<p>Hoo-boy. I see tropes. I see antivaccine tropes. So. Many. Tropes. Also, if you link to Collective Evolution, you lose. It's a wretched hive of scum and quackery. Maybe not as wretched as, say Natural News, but it's pretty bad.</p>
<p>Not surprisingly, it didn't take me long to find this article by antivaccine crank Robert F. Kennedy, Jr., <a href="https://worldmercuryproject.org/news/new-study-vaccine-manufacturers-fda-regulators-used-statistical-gimmicks-hide-risks-hpv-vaccines/" rel="nofollow">Vaccine Manufacturers and FDA Regulators Used Statistical Gimmicks to Hide Risks of HPV Vaccines</a>:</p>
<blockquote><p>
A new study published in <em>Clinical Rheumatology</em> exposes how vaccine manufacturers used phony placebos in clinical trials to conceal a wide range of devastating risks associated with HPV vaccines. Instead of using genuine inert placebos and comparing health impacts over a number of years, as is required for most new drug approvals, Merck and GlaxoSmithKline spiked their placebos with a neurotoxic aluminum adjuvant and cut observation periods to a matter of months.</p>
<p>Researchers from Mexico’s National Institute of Cardiology pored over 28 studies published through January 2017—16 randomized trials and 12 post-marketing case series—pertaining to the three human papillomavirus (HPV) vaccines currently on the market globally. In their July 2017 peer-reviewed <a href="https://www.ncbi.nlm.nih.gov/pubmed/28730271">report</a>, the authors, Manuel Martínez-Lavin and Luis Amezcua-Guerra, uncovered evidence of numerous adverse events, including life-threatening injuries, permanent disabilities, hospitalizations and deaths, reported after vaccination with GlaxoSmithKline’s bivalent Cervarix vaccine and Merck’s quadrivalent or nine-valent HPV vaccines (Gardasil and Gardasil 9). Pharmaceutical company scientists routinely dismissed, minimized or concealed those injuries using statistical gimmicks and <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565961/">invalid comparisons</a> designed to diminish their relative significance.
</p></blockquote>
<p>I note that the article linked to under "<a href="https://scienceblogs.com/www.ncbi.nlm.nih.gov/pmc/articles/PMC3565961/">invalid comparisons</a>" was published by antivaccine activists <a href="http://scienceblogs.com/insolence/2016/03/01/one-more-time-theres-no-evidence-gardasil-causes-premature-ovarian-failure/">Lucija Tomljenovic</a>, Judy Wilyman (yes, that <a href="http://scienceblogs.com/insolence/2016/01/13/the-university-of-wollongong-issues-a-phd-in-antivaccine-pseudoscience/">Judy Wilyman</a>), Eva Vanamee, <a href="http://scienceblogs.com/insolence/2015/10/20/yet-another-antivaccine-rally-yet-another-yawn/">Toni Bark</a>, and <a href="http://scienceblogs.com/insolence/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">Christopher A Shaw</a>. (More on why the comparisons are not invalid later, when I get into the article itself.)</p>
<h2>Roll the tape: A biased, crappy "review" article designed to demonize HPV vaccines</h2>
<p>So let's look at the review article, which is by Manuel Martínez-Lavín and Luis Amezcua-Guerra at the National Institute of Cardiology in Mexico City. So, the first thing I wondered is: WTF is a vaccine article doing being published by <em>Rheumatology</em>? The second thing I wondered was: The National Institute of Cardiology in Mexico? Doing a vaccine study? It certainly is odd. The next thing I wondered was this: Who the heck are Manuel Martínez-Lavín and Luis Amezcua-Guerra? The name Manuel Martínez-Lavín seemed to ring a bell. I seem to recall having heard it before. Fortunately, almighty Google helped out. All I had to do was to search each name with "HPV," "Gardasil," or "Cervarix." I immediately discovered that Martínez-Lavín is not exactly what one would call an...unbiased...source. Let's just put it this way. When you're approvingly cited by the antivaccine website S<a href="http://sanevax.org/hpv-vaccination-syndrome-a-questionnaire-based-study/">aneVax</a>, a site that's <a href="http://scienceblogs.com/insolence/2013/01/09/dr-sin-hang-lee-is-at-it-again-with-the-gardasil-fearmongering/">dedicated</a> to <a href="http://scienceblogs.com/insolence/2012/08/24/despicable-a-parents-guide-to-fishing-for-proof-that-vaccines-kill/">fear mongering about Gardasil</a>. It also turns out that Martínez-Lavín is not above doing highly dubious surveys without controls. From these, I learned that Martínez-Lavín apparently believes that <a href="http://hsionline.com/2015/09/29/hpv-vaccine-3/">HPV vaccines cause fibromyalgia</a>, despite the lack of evidence that it does anything of the sort. Somehow I missed this horrible study, but fortunately <a href="https://www.skepticalraptor.com/skepticalraptorblog.php/hpv-vaccine-anecdotes-not-the-basis-of-real-science/">Skeptical Raptor</a> and <a href="https://drjengunter.wordpress.com/2015/06/01/the-independent-claims-hpv-vaccine-unsafe-science-says-the-independent-is-wrong/">Dr. Jen Gunter</a> did not. Now, I must realize that I didn't find much about Luis Amezcua-Guerra, but it's not hard to see how biased the article by Manuel Martínez-Lavín and Luis Amezcua-Guerra is, even before looking at it in detail.</p>
<p>Let's look at the first trope, which is a common anti-Gardasil trope that it is an inappropriate control to compare an aluminum-containing vaccine like Gardasil to an aluminum adjuvant without the actual antigens from the vaccine. The argument is that the best control should have been normal saline; i.e., an inert control. This is a profoundly ignorant argument when you have an intervention known to be safe based on many studies in many vaccines over the years (like aluminum adjuvants). When you have such an ingredient, then if you want to determine whether or not a vaccine containing that ingredient works and is safe, an excellent way to do it is to compare it to a control containing everything in the vaccine except the antigens that produce the immune response. In other words, the adjuvant-only control is a very good control. Channeling antivaccine tropes aplenty, the authors of the review try their best to convince you that the real reason this control was chosen in so many studies of Gardasil and Cervarix was to hide adverse events due to these vaccines. Of course, the existence of long term studies (like <a href="http://www.skepticalraptor.com/skepticalraptorblog.php/gardasils-safety-effectiveness-part-47-long-term-study/">this one</a>) comparing HPV vaccines to saline placebo controls rather undermines this particular antivaccine talking point. Basically, we have evidence from both studies comparing HPV vaccines to adjuvant-only controls and to saline controls showing that HPV is both effective and safe.</p>
<p>Again, go back to Lucija Tomljenovic's paper cited earlier. That's exactly the main claim in the paper. Let's just put it this way, if an antivaccine crank like her makes an argument, that's a pretty good indication that' what's being argued is pure, grade A BS. Of course, this is pure, grade A BS independent of Tomljenovic's use of it. A good way of looking at it is that Tomljenovic uses it because she is an antivaxers and antivaxers gravitate to claims about vaccines that are grade A unadulterated BS. Also look at it this way: If Martínez-Lavín and Amezcua-Guerra think this is a such a compelling argument that they make it the centerpiece of their "analysis" of the randomized controlled trials (RCT) of Gardasil and Cervarix, it starts to make me question everything else in the paper. Adding to that impression is my perusal of the references, which include the works of such antivaccine "scientists" as, yes, Tomljenovic, Yehuda Shoenfeld (who just <a href="http://retractionwatch.com/2017/08/15/paper-extremely-flawed-journal-retracts-paper-heavy-criticism/">had a paper retracted</a>), Deirdre Therese Little (who is on the board of advisors of an conservative Catholic Australian antivaccine group that preaches that Gardasil leads to promiscuity and who <a href="http://scienceblogs.com/insolence/2016/03/01/one-more-time-theres-no-evidence-gardasil-causes-premature-ovarian-failure/">promotes the false message</a> that Gardasil causes premature ovarian failure). Indeed, the paper by Little cited was <a href="http://scienceblogs.com/insolence/2012/10/30/ovarian-failure-caused-by-gardasil-not-so-fast/">deconstructed by yours truly</a> when it came out.</p>
<p>Not surprisingly, this isn't even a very good systematic or "critical" review. It's definitely not a meta-analysis, although our intrepid authors do try to make it sound like one by "reanalyzing" data from the papers they want to try to refute. Most of them didn't show any difference in adverse events in placebo or HPV vaccine group. There were two, however, that, according to the authors, did:</p>
<blockquote><p>
Two of the largest HPV vaccine randomized trials did find significantly more severe adverse events in the tested vaccine group vs. the comparator group: The 4-year interim follow-up VIVIANE study safety analysis compared 2881 healthy women older than 25 years injected with the bivalent HPV vaccine vs. 2871 age-matched women injected with aluminum placebo [29]. As expected in large randomized trials, both groups displayed remarkably similar baseline characteristics. General solicited symptoms during the 7-day post-vaccination period occurred more often in the HPV vaccine group (65%) than those in the control group (58%). Our calculated 2 × 2 contingency table p value was <0.01. Vaccine-related general solicited symptoms during the 7-day post-vaccination period were also more frequent after HPV vaccination (41%) than those after placebo injection (36%) p </p></blockquote>
<p>This is just plain silly. The authors are doing 2x2 contingency tables because they didn't have the raw data, while the authors of the original VIVIANE study did and did much more sophisticated analyses. Here's what the the <a href="http://www.thelancet.com/journals/lancet/article/PIIS0140-6736(14)60920-X/abstract">actual VIVIANE study</a> says about this:</p>
<blockquote><p>
Solicited injection-site symptoms occurred in more women in the vaccine group than in the control group (table 4). General solicited symptoms during the 7-day post-vaccination period occurred slightly more often in the vaccine group than in the control group. The incidence of unsolicited symptoms, serious adverse events, medically significant conditions, new-onset chronic disease, and new-onset autoimmune disease was similar in both groups, and pregnancy outcomes did not differ between groups (table 4). 17 deaths occurred, 14 (<1%) of 2881 women in the vaccine group and three (<1%) of 2871 in the control group; none of the deaths were believed to be related to vaccination. The independent data monitoring committee did an unblinded review of all deaths; the causes of death were very variable and no cluster of disease type was noted (appendix p 4). The mean time between the last vaccination and death was 682 days (SD 321) in the vaccine group and 496 days (424) in the control group (range 67–1191 days for both groups), suggesting no temporal relation between vaccination and death.
</p></blockquote>
<p>In other words, the difference was mainly more injection site problems, which would be expected for an active vaccine being compared to just the adjuvant. One would expect more inflammatory reactions. As for the deaths, they were analyzed by an independent data monitoring committee and showed no pattern that suggested a link to the vaccine. Taking a look at the list in the appendix should tell you that it's unlikely there was a link (click to embiggen):</p>
<p><a href="https://scienceblogs.com/files/insolence/files/2017/08/GardasilDeaths.png"><img src="http://scienceblogs.com/insolence/files/2017/08/GardasilDeaths-450x287.png" alt="" width="450" height="287" class="aligncenter size-medium wp-image-11002" /></a></p>
<p>For instance, there are three suicides, one homicide (it strains credulity, even for antivaxers, to think that HPV vaccination can cause one to become a murder victim), two cases of breast cancer (both women were in their late 40s), a case of drug hypersensitivity and renal failure. You get the idea.</p>
<p>The authors look at another <a href="http://www.nejm.org/doi/full/10.1056/NEJMoa1405044">randomized double blind RCT</a> of Gardasil that contrasted the 9-valent dose versus the quadrivalent formulation. Basically, instead of four serotypes, there are nine in the newer vaccine. I'm fed up with this "review" already; so I'm just going to cite the actual RCT:</p>
<blockquote><p>
The recipients of the 9vHPV vaccine were more likely than the recipients of the qHPV vaccine to have adverse events related to the injection site (90.7% vs. 84.9%), with the most common events (incidence ≥2%) being pain, swelling, erythema, and pruritus (Table Adverse Events.); more than 90% of these events were mild to moderate in intensity. Events of severe intensity were more common in the 9vHPV group. The frequency of systemic adverse events was generally similar in the two groups — 55.8% in the 9vHPV vaccine group and 54.9% in the qHPV vaccine group. The most common systemic adverse events related to vaccination (incidence ≥2%) were headache, pyrexia, nausea, dizziness, and fatigue. Less than 0.1% of participants discontinued study vaccination because of a vaccine-related adverse event. All the serious adverse events are listed according to system organ class in Tables S6 and S7 in the Supplementary Appendix. Pregnancy was reported in 1192 participants in the 9vHPV group and 1129 participants in the qHPV group, and information on outcomes was available for approximately 85% of these pregnancies (Table S8 in the Supplementary Appendix). The proportions of participants with live births, difficulty with delivery, spontaneous abortions, and late fetal deaths were similar in the two groups. Congenital anomalies were reported in a total of 32 infants and 9 fetuses (20 in the 9vHPV group and 21 in the qHPV group). No congenital anomaly was reported in the case of pregnancies with an estimated date of conception that was within 30 days before or after any vaccination (these pregnancies represented approximately 8% of the total number of pregnancies with known outcomes).
</p></blockquote>
<p>So basically, there were more injection site problems in the 9vHPV group, which is not surprising for a more immunogenic vaccine, and there were systemic symptoms like headache, pyrexia, nausea, dizziness, and fatigue, but clearly not that bad if only 0.1% of the participants stopped the three dose series as a result. Not understanding the concept of number needed to treat with respect to vaccines, the authors plunge boldly ahead:</p>
<blockquote><p>
The average number needed to vaccinate with the 9- valent dose to prevent one episode of the pre-specified primary end-points that would not otherwise have been prevented by the 4-valent immunization is 1757 with 95% CI ranging from 131 to infinity
</p></blockquote>
<p>That's actually pretty damned good for a vaccine compared to another. If you vaccinate millions of women the advantage of the 9vHPV vaccine would become apparent in the form of thousands of additional cases of HPV prevented. Also remember that the 4vHPV contains the four most common types of HPV associated with cervical cancer. Adding five more types would, by definition, add less common types of HPV and produce less benefit.</p>
<h2>And the "cover-up" continues</h2>
<p>Finally, the authors think that the post-marketing studies are "covering up" adverse events from HPV vaccines. In their table (Table 2), they cite a whole pubnch of studies, including the one by Martínez-Lavin that I cited above that was nothing more than a questionnaire-based study in which he claims to have found an association between HPV vaccination and fibromyalgia in 45 women. They also cite—you guessed it—Tomljenovic twice, including three papers claiming to find the "ASIA syndrome" post-vaccination. It's a syndrome <a href="http://scienceblogs.com/insolence/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">so vaguely defined</a> and (of course) not accepted by anyone other than Yehuda Shoenfeld and his fellow travelers as a legitimate medical syndrome. They also include dubious papers claiming to link Gardasil to premature ovarian failure.</p>
<p>The authors then go on to cite a number of postmarketing studies looking at HPV vaccine adverse events. A lot of these studies found increased incidence of syncopal symptoms (like dizziness and occasionally passing out) after vaccination. Of course, that's why we insist that children receiving any vaccination, but Gardasil in particular (given the propensity of girls of the age receive Gardasil have syncopal episodes after blood draws and injections anyway), be observed for a while after receiving the dose. It's not a reason not to vaccinate. Injections of all types are associated with fainting, as the authors of several of the papers note and the authors of this review do not.</p>
<p>A <a href="https://www.ncbi.nlm.nih.gov/pubmed/26107345">much better review of postlicensure studies of HPV vaccines</a> by Vichnin et al found that only "syncope, and possibly skin infections were associated with vaccination in the postlicensure setting" and that serious adverse events, "such as adverse pregnancy outcomes, autoimmune diseases (including Guillain-Barre Syndrome and multiple sclerosis), anaphylaxis, venous thromboembolism and stroke, were extensively studied, and no increase in the incidence of these events was found compared with background rates."</p>
<p>Overall, this is a terrible systematic review. It is clearly designed to make HPV vaccination look as bad as the authors can make it look by playing up known adverse events due to HPV vaccines, such as syncope, claiming that adverse events are vastly underreported, and citing papers by antivaccine cranks as "evidence" that there are all sorts of horrible things caused by Gardasil that "They" don't want you to know about. Not surprisingly, it's spreading in the antivaccine crankosphere. Surprisingly, I haven't seen it on Natural News yet. It's coming, though. I'm sure of it.</p>
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Wed, 16 Aug 2017 07:40:58 +0000oracknows22606 at https://scienceblogs.comThe crank medical organization to which HHS nominee Dr. Tom Price belongs lays down a heaping helping of antivaccine pseudosciencehttps://scienceblogs.com/insolence/2016/12/22/the-crank-medical-organization-to-which-hhs-nominee-dr-tom-price-belongs-lays-down-a-heaping-helping-of-antivaccine-pseudoscience
<span>The crank medical organization to which HHS nominee Dr. Tom Price belongs lays down a heaping helping of antivaccine pseudoscience</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>Three weeks ago, I wrote a post that, much to my surprise, went viral, garnering more Facebook "Likes" than any before it, although it only came in maybe third in traffic after the all-time record-holding post from a couple of years ago. Maybe I shouldn't have been surprised. It was, after all, about Tom Price, Donald Trump's nominee for Secretary of the Department of Health and Human Services (HHS). What I noted that apparently caught the attention of many times more people than my usual daily brain droppings usually do was that Tom Price belongs to <a href="http://scienceblogs.com/insolence/2016/11/30/the-new-secretary-of-health-and-human-services-is-a-member-of-a-fringe-medical-organization-heres-what-that-means/">one of the wingnuttiest of wingnut medical groups</a> I've ever encountered, the Association of American Physicians and Surgeons (AAPS). There's no need for me here to reiterate the group's Ayn Rand-like worship of the brave maverick doctor <em>über alles</em> in detail, as I've done that before multiple times <a href="http://scienceblogs.com/insolence/2006/03/14/journal-of-american-physicians/">beginning nearly eleven years ago</a>. Basically, to the AAPS, doctors should not accept payment from Medicare (which it views as unconstitutional) and that the autonomy of doctors, who are portrayed as akin to <a href="https://en.wikipedia.org/wiki/John_Galt">John Gal</a>t and other "producers," should never be constrained by pesky, puny things like evidence-based guidelines, because, apparently, every doctor is expert enough to interpret the vast medical literature without any help.</p>
<p>On the other hand, it is worth briefly mentioning the pure antiscience and pseudoscience that emanates from the AAPS, particularly through its house organ, the <a href="http://www.jpands.org" rel="nofollow">Journal of American Physicians and Scientists</a> (JPANDS). This journal is a veritable cornucopia of ideology-motivated quackery and pseudoscience, including antivaccine pseudoscience up to and including the despicable claim that shaken baby syndrome is a "<a href="http://scienceblogs.com/insolence/2006/03/14/journal-of-american-physicians/">misdiagnosis for mercury poisoning</a>" and that <a href="http://jpands.org/hacienda/article25.html" rel="nofollow">sudden infant death syndrome might be caused by vaccines</a>, the <a href="http://scienceblogs.com/insolence/2007/10/25/abortion-and-breast-cancer-the-chicago-t/">bogus claim that abortion causes breast cancer</a>, and anthropogenic global climate change denialism (don't ask what that's doing there). Indeed, Dr. Jane Orient, the executive director of AAPS, <a href="https://sciencebasedmedicine.org/hostility-towards-scientific-consensus-a-red-flag-identifying-a-crank-or-quack/">denies the very concept of a scientific consensus</a>.</p>
<!--more--><p>Well, the <a href="http://jpands.org/jpands2104.htm" rel="nofollow">Winter 2016 issue of JPANDS</a> is out, hot off the presses, digitally speaking, and I can't help but wonder: What does Dr. Price think of some of the articles found therein. In particular, I'm interested in what he thinks of an article by Neil Z. Miller entitled <a href="http://www.jpands.org/vol21no4/miller.pdf" rel="nofollow">Aluminum in Childhood Vaccines Is Unsafe</a>. Well, you have to say one thing for Miller: He's consistent. Just two issues earlier, Miller published an article in JPANDS entitled <a href="http://scienceblogs.com/insolence/2016/11/30/the-new-secretary-of-health-and-human-services-is-a-member-of-a-fringe-medical-organization-heres-what-that-means/" rel="nofollow">Combining Childhood Vaccines at One Visit Is Not Safe</a>. I'm eagerly looking forward to an ongoing series from Miller: <em>MMR Is Not Safe</em>. <em>Thimerosal Is Not Safe</em>. <em>All Those Nasty Toxins in Vaccines Are Not Safe</em>. Not surprisingly, I took notice of Miller's last article in JPANDS and <a href="http://scienceblogs.com/insolence/2016/06/16/combining-childhood-vaccines-at-one-visit-is-not-safe-not-so/">applied a not-so-Respectful Insolence</a> to misinformation and pseudoscience that deserved much worse. Even less surprisingly, once having seen Miller's second JPANDS publication, I can't resist a repeat, particularly now that I know that Dr. Price is a member in good standing of the AAPS. These are the sorts of misinformation-packed articles that need to be thrown into Price's face during his confirmation hearings in order to force him to justify why he belongs to an organization so opposed to accepted medical science.</p>
<p>So let's take a look, shall we?</p>
<p>Not content with just demonizing aluminum, which is used as an adjuvant for some vaccines. Basically, an adjuvant is a substance that, when injected with vaccines, can result in a more intense immune response. Miller has to start with the dreaded mercury-containing preservative thimerosal. There's a method to his madness, of course. It's not a method that makes any scientific sense, but rather is designed to draw attention from the very simple observation that, since the phase-out of thimerosal as a preservative in vaccines in the US, autism prevalence hasn't declined. Quite the contrary, actually. Miller opines:</p>
<blockquote><p>
From 1999 through 2002, several vaccines containing mercury were phased out of the childhood immunization schedule. Manufacturing of childhood vaccines with thimerosal ceased in 2001, but those that were not past their expiration date remained on the market for sale until January 2003.<sup>1</sup> They were replaced with low-mercury or “thimerosal-free” vaccines. In the years that followed, autism rates continued to rise, prompting health authorities to assert that autism is not linked to mercury in vaccines and that vaccination policies are safe and appropriate.<sup>2-4</sup> (If mercury in vaccines contributed to autism, then rates should have dropped after mercury was removed.) However, in 2002, during this so-called phase-out period, the Centers for Disease Control and Prevention (CDC) actually added two doses of mercury-containing influenza vaccines to the list of inoculations urged for all babies 6 to 23 months of age.<sup>5</sup> Two years later, the CDC also added pregnant women in their first trimester to the list of people officially recommended and actively encouraged to receive influenza vaccines, even though a majority of available doses contained mercury.<sup>6</sup></p>
<p>In addition to these questionable actions during this highly publicized “phase-out” of mercury, four doses of a new vaccine with high aluminum content were added to the childhood immunization schedule in February 2000 (for pneumococcus) and two doses of another aluminum-containing vaccine (for hepatitis A) were added in 2005.<sup>7,8</sup> These changes to the vaccine schedule resulted in a substantial increase of aluminum-containing vaccine doses—from 10 to 16 injections—that babies are still mandated to receive by 18 months of age.
</p></blockquote>
<p>Notice what Miller left out? Well, he left out multiple things. However, the most glaring is a simple matter of quantity. How much mercury was in the childhood vaccination schedule after 2003 compared to before? The answer, of course, is a lot less, even with the addition of the new vaccines. For instance, in an Italian study from the 1990s testing the existing DTaP vaccine versus a then-new thimerosal-free version, children received 137.5 μg of mercury and just eliminating the thimerosal from the DTaP in that schedule <a href="http://scienceblogs.com/insolence/2009/01/27/the-first-of-i-hope-many-very-bad-days-f/">cut mercury exposure by more than half</a>. Moreover, while it's true that most flu vaccines then still contained thimerosal, it didn't take long for manufacturers to get rid of the thimerosal, particularly in the childhood vaccines. These days, it's so hard to find thimerosal-containing flu vaccines that when I get my yearly dose, I often joke about asking to add extra thimerosal.</p>
<p>Of course, the narrative that Miller is selling is that the reason autism prevalence didn't plunge a few years after the removal of nearly all the thimerosal from the childhood vaccine schedule by early 2003 is because of an increase in aluminum exposure. This idea is as much a pile of nonsense as the idea that thimerosal was responsible for an "autism epidemic," not the least of which because it would be an incredibly coincidence that, if you accept the rationale of someone like Miller that both thimerosal and aluminum can contribute to autism, just as one autism-containing vaccine ingredient was removed another was added in sufficient quantity to cause autism prevalence to keep climbing at the very same rate that it was climbing before. That's because antivaccinationists have always known that if autism prevalence kept climbing after thimerosal was removed from vaccines it would be a deadly blow against their <a href="http://scienceblogs.com/insolence/2010/09/17/safeminds-swings-at-price-et-al-and-miss/">belief that thimerosal causes autism</a> when <a href="http://scienceblogs.com/insolence/2010/09/14/price-et-al-roundup-blaming-mercury-in-v/">it doesn't</a>. So basically, Miller tries to argue that a 25% increase in aluminum exposure due to vaccines was enough to make up for—scratch that, more than make up for—the loss of thimerosal in its claimed evil autism-causing properties.</p>
<p>To demonstrate the "toxicity" of aluminum, Mr. Miller has to do some rather major contortions, so much so that it looks very much as though he's playing Twister with vaccine science, and the construct can't stand:</p>
<blockquote><p>
Aluminum is neurotoxic and has a long history of well-documented hazards.<sup>14</sup> For example, as early as 1921 The Lancet described a 46-year-old metal worker in whom “aluminium produced a rather slow intoxication. In this case it caused memory loss, tremor, jerky movements and incontinence of urine.”<sup>15</sup> In 1927, Dr. Victor Vaughn, a toxicologist with the University of Michigan, testified before the Federal Trade Commission that “all salts of aluminum are poisonous when injected subcutaneously or intravenously.”<sup>16</sup> By 1951, Chusid et al. showed that chronic epilepsy could be induced in monkeys through intra-cerebral administration of aluminum hydroxide cream.<sup>17</sup> In 1968, Driver et al. performed a similar experiment by placing aluminum hydroxide cream unilaterally on the posterior parietal cortex of six monkeys.<sup>18</sup> From 3 to 8 weeks after surgery, electrical abnormalities could be seen on an electroencephalogram and the monkeys exhibited “episodic twitching of the limbs and face.” The animals were also impaired at learning new tasks and at re-learning tasks first learned prior to the intervention.
</p></blockquote>
<p>Aluminum exposure in a metal worker in the 1920s was due to a much larger exposure than any vaccine or series of vaccines would be expected to produce. Slathering large quantities of aluminum onto the cerebral cortex is much different than injecting tiny quantities into the muscle. Dose and poison, Mr. Miller. Dose and poison. Learn it, live it, love it. The dose makes the poison, and the dose of <a href="http://www.fda.gov/BiologicsBloodVaccines/ScienceResearch/ucm284520.htm">aluminum in vaccines is quite safe</a>. I also can't help but notice some of the articles referenced by Miller are not exactly from reliable sources. For instance, reference #16 is from David Ayoub's talk to the National Autism Association Conference in 2007. That's a serious antivaccine conference, and David Ayoub is know for his conspiratorial thinking and belief in <a href="https://leftbrainrightbrain.co.uk/2006/08/18/david-ayoub-black-helicopters-and-social-movement/">black helicopters and the Illuminati</a>. It's not for nothing that Ayoub was <a href="http://americanloons.blogspot.com/2013/03/462-david-ayoub.html">inducted into the Encylopedia of American Loons</a>. If you don't believe me, just look for yourself at this typical talk by Ayoub:</p>
<iframe width="560" height="315" src="https://www.youtube.com/embed/S39kmstBNkk" frameborder="0" allowfullscreen=""></iframe><p>
That's some weapons-grade crazy.</p>
<p>Mr. Miller also liberally cites Lucija Tomljenovic and Christoper Shaw, two antivaccine cranks par excellence whose abuse of science has been discussed many times <a href="http://scienceblogs.com/insolence/?s=%22Lucija+Tomljenovic%22+OR+%22Christopher+Shaw%22+vaccines">here</a> and <a href="http://www.skepticalraptor.com/skepticalraptorblog.php/tag/lucija-tomljenovic/">elsewhere</a>. Let's just put it this way: Tomljenovic and Shaw very much believe that Gardasil causes premature ovarian failure (it doesn't, and <a href="http://scienceblogs.com/insolence/2016/03/01/one-more-time-theres-no-evidence-gardasil-causes-premature-ovarian-failure/">here's why</a>) and that it kills (it doesn't, and <a href="http://scienceblogs.com/insolence/2012/10/31/and-now-death-by-gardasil-again-not-so-fast/">here's why</a>). Mr. Miller even cites one of the most hilariously misbegotten articles blaming aluminum in vaccines for autism, one that so hilariously confuses correlation with causation that it makes blaming global warming on the decrease in number of pirates <a href="http://scienceblogs.com/insolence/2011/12/08/and-global-warming-is-caused-by-the-decr/">seem reasonable by comparison</a>. Yes, it's a Tomljenovic and Shaw paper.</p>
<p>The rest of the paper basically defines the term "cherry picked." Mr. Miller lists, in maximally frightening terms, a series of studies that purport to find significant negative health consequences (as opposed, for instance, to injection site pain or complications, something common to all vaccines, regardless of whether they contain aluminum or not), often by antivaccine cranks, and unsurprisingly ignores the <a href="http://www.skepticalraptor.com/skepticalraptorblog.php/aluminum-adjuvant-vaccines-cherry-picking/">vast existing literature on the safety of aluminum adjuvants</a>. These studies often ignore the principle of "the dose makes the poison":</p>
<blockquote><p>
According to the American Academy of Pediatrics (AAP), “Aluminum is now being implicated as interfering with a variety of cellular and metabolic processes in the nervous system and in other tissues.”<sup>19</sup> Bishop et al. published data showing that “aluminum accumulates in the body when protective gastrointestinal mechanisms are bypassed, renal function is impaired, and exposure is high.”<sup>20</sup> For example, in premature infants, “prolonged intravenous feeding with solutions containing aluminum is associated with impaired neurologic development” by 18 months of age. More recently, Kawahara et al. published research confirming that “aluminum can cause severe health problems in particular populations, including infants.”<sup>21</sup> The authors of this paper also declared that “whilst being environmentally abundant, aluminum is not essential for life. On the contrary, aluminum is a widely recognized neurotoxin that inhibits more than 200 biologically important functions and causes various adverse effects in plants, animals, and humans.”
</p></blockquote>
<p>Bloody hell. He's quoting the data for aluminum toxicity in premature infants who have prolonged exposure to aluminum in their parenteral (intravenous) nutrition. We're talking about premature babies who are either too premature or too ill to be fed using their guts and as a consequence have to be fed intravenously and thus receive a lot of aluminum. Comparing this exposure to aluminum to exposure due to vaccines is akin to comparing Mt. Everest to the hill near your house that makes you tired if when you have to climb it.</p>
<p>Unbelievably, Mr. Miller even invokes a favorite antivaccine trope used for any scary-sounding ingredient in vaccines, elaborating on the "injection" aspect of vaccines above:</p>
<blockquote><p>
Moreover, vaccines with aluminum adjuvants are injected into the body, bypassing protective barriers of the gastrointestinal tract and skin. Absorption of aluminum by this mode is more efficient than through ingestion, increasing the likelihood of a toxic outcome. The authors summarized their findings: “Evidence has now emerged showing that autism may in part result from early-life immune insults induced by environmental xenobiotics. One of the most common xenobiotic with immuno-stimulating as well as neurotoxic properties to which infants under two years of age are routinely exposed worldwide is the aluminum vaccine adjuvant.”
</p></blockquote>
<p>At least he refrained from the more common variant of this trope, that vaccines are "injected directly into the bloodstream," when in fact they are usually injected intramuscularly. are talking about small amounts of aluminum injected into the muscle, where they are slowly abosrbed.</p>
<p>I could go on and on. Mr. Miller's paper is what we in the skeptic biz call a "target-rich environment." The reason I brought it up now to discuss has less to do with the misinformation in the article, of which there is plenty, or its unusualness, of which there is none. Rather, it's because, now, less than three weeks after Dr. Price was nominated to head HHS, the organization to which he belongs is still at it, publishing antivaccine propaganda every bit as egregious as it's been doing at least since 2006. I don't know about you, but I want to know how many beliefs Dr. Price shares with the organization to which he belongs. I want to see him have to read this article and explain himself, specifically, whether he shares the AAPS's antivaccine views.</p>
<p>Yes, there are a lot of other areas where Dr. Price's views are very worrisome, but this is one that shouldn't be ignored.</p>
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<span><a title="View user profile." href="https://scienceblogs.com/author/oracknows">oracknows</a></span>
<span>Wed, 12/21/2016 - 21:15</span>
Thu, 22 Dec 2016 02:15:01 +0000oracknows22456 at https://scienceblogs.comNo, Gardasil does not cause behavioral problemshttps://scienceblogs.com/insolence/2016/02/17/no-gardasil-does-not-cause-behavioral-problems
<span>No, Gardasil does not cause behavioral problems</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>Believe it or not, I frequently peruse <a href="http://retractionwatch.com">Retraction Watch</a>, the blog that does basically what its title says: It watches for retracted articles in the peer-reviewed scientific literature and reports on them. Rare is it that a retracted paper gets by the watchful eyes of the bloggers there. So it was that the other day I noticed an post entitled <a href="http://retractionwatch.com/2016/02/15/journal-temporarily-removes-paper-linking-hpv-vaccine-to-behavioral-issues/">Journal temporarily removes paper linking HPV vaccine to behavioral issues</a>. I noticed it mainly because it involves a paper by two antivaccine "researchers" whom we've met several times before, Christopher A. Shaw and Lucija Tomljenovic in the Department of Ophthalmology at the University of British Columbia. Both have a long history of publishing antivaccine "research," mainly falsely blaming the aluminum adjuvants in vaccines for autism and, well, just about any health problem children have and blaming Gardasil for <a href="http://scienceblogs.com/insolence/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">premature ovarian failure</a> and all manner of woes <a href="http://scienceblogs.com/insolence/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">up to and including death</a>. Shaw even prominently featured in the rabidly antivaccine movie <a href="http://scienceblogs.com/insolence/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">The Greater Good</a>.</p>
<p>Normally, Shaw and Tomljenovic tend to publish their antivaccine spew in bottom-feeding journals, but what also caught my attention was that this time they seemed to have managed to score a paper in a journal with a good reputation and a <a href="https://en.wikipedia.org/wiki/Vaccine_(journal)">reasonable impact factor</a>: <a href="http://www.journals.elsevier.com/vaccine">Vaccine</a>. Here's the story from <a href="http://retractionwatch.com/2016/02/15/journal-temporarily-removes-paper-linking-hpv-vaccine-to-behavioral-issues/">Retraction Watch</a>:</p>
<!--more--><blockquote>
The editor in chief of <em>Vaccine</em> has removed a paper suggesting a human papillomavirus (HPV) vaccine can trigger behavioral changes in mice.
<p>The note doesn’t provide any reason for the withdrawal, although authors were told the editor asked for further review.</p>
<p>Two co-authors on the paper — about Gardasil, a vaccine against HPV — have previously suggested that aluminum in vaccines is linked to autism, in research a World Health Organization advisory body concluded was “seriously flawed.”</p>
<p>Approximately 80 million doses of Gardasil were administered in the U.S. between 2006 and 2015. Both the the WHO and the Centers for Disease Control and Prevention have ruled the vaccine to be safe — the CDC, for instance, calls it “safe, effective, and recommended.”</p>
<p>The journal published an uncorrected proof of “Behavioral abnormalities in young female mice following administration of aluminum adjuvants and the human papillomavirus (HPV) vaccine Gardasil” online on January 9th, 2016. In its place now is a note that says:</p>
<blockquote><p>
The publisher regrets that this article has been temporarily removed. A replacement will appear as soon as possible in which the reason for the removal of the article will be specified, or the article will be reinstated.
</p></blockquote>
<p>Since the article had not yet been officially published in the journal, it’s not indexed by Thomson Scientific’s Web of Knowledge.
</p></blockquote>
<p>Curiouser and curiouser. Fortunately for me, an online bud happened to have downloaded the paper and supplied me with the PDF. I say this because, as noted above, the paper is <a href="http://www.ncbi.nlm.nih.gov/pubmed/26778424">no longer available</a> on <a href="http://www.sciencedirect.com/science/article/pii/S0264410X16000165">the <em>Vaccine</em> website</a>. Its corresponding author, it turns out, is Yehuda Shoenfeld, <a href="http://scienceblogs.com/insolence/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">whom we have also met before</a> and have encountered speaking at antivaccine conferences. In antivaccine circles, Shoenfeld is best known for coining the term "ASIA" ("Autoimmune/Inflammatory Syndrome Induced by Adjuvants"). Did I say "coin the term"? Really, I should have said "pulled the term out of his nether regions, leaving a coating of what one's nether regions generally expels all over it." Because ASIA is a made-up syndrome with no compelling evidence that it's real. Basically, as <a href="http://scienceblogs.com/insolence/2013/08/09/antivaccinationists-against-the-hpv-vaccine-round-5000/">I've described before</a>, its criteria are so vague as to be able to be applied to almost anything.</p>
<p>Oh, and he <a href="http://web.archive.org/web/20110514140345/http://www.neurodiversity.com/weblog/article/136/">edited a journal</a> very sympathetic to antivaccine "science."</p>
<p>So right off the bat, I knew something fishy was going on here, and, thanks to the person who supplied me with a PDF of the article, I knew I had to take a look to see if I could figure out what happened. What I can say, having read the article, is that it is so shoddily done that it represents a massive failure of peer review that a journal as good as <em>Vaccine</em> ever accepted it for publication. I don't know who the editor to whom this manuscript was assigned was (clearly it wasn't <em>Vaccine</em>'s editor-in-chief Gregory Poland, as it was he who requested temporary removal of the paper), but whoever it was should hang his or her head in shame and resign in disgrace from the editorial board of the journal.</p>
<p>I really have to wonder if this travesty is the result of the problem of "fake peer review. This is a problem that has come to light recently that takes advantage of the practice of some journals to allow authors to suggest peer reviewers for their manuscript. I perused <em>Vaccine</em>'s Instructions for Authors and found that the journal encourages authors to suggest up to five potential reviewers. I've <a href="http://scienceblogs.com/insolence/2015/12/28/fixing-peer-review/">described the problem before</a>, and Vaccine editors ought to stop this practice immediately. One can't help but wonder who reviewed this manuscript. One almost has to wonder if it was Andrew Wakefield, given how bad the manuscript is.</p>
<p>I'll show you what I mean. <a href="http://www.skepticalraptor.com/skepticalraptorblog.php/gardasil-causes-behavioral-issues-more-myth-debunking/">Skeptical Raptor</a> has already discussed this study. He's also done an excellent job of laying out the safety studies involving millions of doses of Gardasil that show that Gardasil is incredibly safe. His post is almost enough. Almost. However, in this discussion, I have the advantage of having the actual PDF of the paper in front of me. As good as the Raptor is, he lacked that.</p>
<p>Since the paper isn't currently available, and I'm not about to violate copyright by making the whole PDF available to anyone who wants it, let's look at the abstract, and then I'll discuss the actual paper's findings, such as they are:</p>
<blockquote><p>
Vaccine adjuvants and vaccines may induce autoimmune and inflammatory manifestations in susceptible individuals. To date most human vaccine trials utilize aluminum (Al) adjuvants as placebos despite much evidence showing that Al in vaccine-relevant exposures can be toxic to humans and animals. We sought to evaluate the effects of Al adjuvant and the HPV vaccine Gardasil versus the true placeboon behavioral and inflammatory parameters in young female mice. Six week old C57BL/6 female micewere injected with either, Gardasil, Gardasil + pertussis toxin (Pt), Al hydroxide, or, vehicle control inamounts equivalent to human exposure. At six months of age, Gardasil and Al-injected mice spent sig-nificantly more time floating in the forced swimming test (FST) in comparison to vehicle-injected mice(Al, p = 0.009; Gardasil, p = 0.025; Gardasil + Pt, p = 0.005). The increase in floating time was already highlysignificant at three months of age for the Gardasil and Gardasil + Pt group (p ≤ 0.0001). No significant differences were observed in the number of stairs climbed in the staircase test nor in rotarod performance,both of which measure locomotor activity. Since rotarod also measures muscular strength, collectivelythese results indicate that differences observed in the FST were not due to locomotor dysfunction, butlikely due to depression. Additionally, at three months of age, compared to control mice, Al-injectedmice showed a significantly decreased preference for the new arm in the Y maze test (p = 0.03), indi-cating short-term memory impairment. Moreover, anti-HPV antibodies from the sera of Gardasil andGardasil + Pt-injected mice showed cross-reactivity with the mouse brain protein extract. Immunohisto-chemistry analysis revealed microglial activation in the CA1 area of the hippocampus of Gardasil-injectedmice compared to the control. It appears that Gardasil via its Al adjuvant and HPV antigens has the abilityto trigger neuroinflammation and autoimmune reactions, further leading to behavioral changes.
</p></blockquote>
<p>I'm quite familiar with C57BL/6 mice. Indeed, I cut my research teeth, so to speak, studying mouse tumor models and the induction of angiogenesis (new blood vessel growth) back in the late 1990s. Now, right from the beginning, I have to wonder who the peer reviewers were to have accepted statements like those in the very first few sentences; i.e., claims that vaccine adjuvants and vaccines can cause autoimmune disease (basically the only people claiming adjuvants can cause autoimmunity are Shoenfeld and antivaccine "scientists" associated with him), that aluminum adjuvants are toxic to humans at vaccine-relevant doses (<a href="http://www.chop.edu/centers-programs/vaccine-education-center/vaccine-ingredients/aluminum#.VsPWXcePbYA">no</a>, <a href="http://www.fda.gov/BiologicsBloodVaccines/ScienceResearch/ucm284520.htm">they aren't</a>).</p>
<p>So what did the authors do? They basically tested whether Gardasil or its aluminum adjuvant cause behavioral issues in mice. Upon perusing the abstract, my first question was this: Why this hypothesis? Why do the authors think that HPV vaccine might cause behavioral changes? We don't really learn why. Their reasoning seems to be "Because vaccines are bad" or "because aluminum is bad." Certainly they don't present any publications with convincing evidence supporting a potential link between vaccines or aluminum adjuvants and behavioral problems. Now, if all this study were doing were testing vaccines or adjuvants in cultured cells, this wouldn't be such a big deal. However, if you're going to subject animals to pain and distress and then kill them at the end to look at their brains, you need some compelling evidence to show (1) why your hypothesis is so compelling and (2) why only an animal model can answer the question you want to answer. Shoenfeld's group utterly fails at this. I can only speculate that animal research regulations must be more lax in Israel than they are in the US.</p>
<p>So let's look at the tests these poor mice were subjected to:</p>
<ul><li><strong>The forced swimming test</strong> (FST): This is a test purported to model depression. How one models depression in a mouse, I don't know, but there is literature to support this use. In any case, increased floating time (not swimming) is indicative of depressive behavior and can also indicate locomotor dysfunction. In these experiments: "mice were placed in individual glass beakers with water 15 cm deep at 25° C. On the first day, mice were placed in the cylinder for a pretest session of 10 min,and later were removed from the cylinder, and then returned totheir home cages. Twenty-four hours later (day 2), the mice were subjected to a test session for 6 min. The behavioral measure scored was the duration (in seconds) of immobility or floating, defined as the absence of escape-oriented behaviors, such as swimming,jumping, rearing, sniffing or diving, recorded during the 6 min test."</li>
<li><strong>Staircase test</strong>: This test is supposed to measure anxiety, as more anxious mice won't explore as much. In these experiments: "The staircase mazeconsisted of a polyvinyl chloride enclosure with five identicalsteps, 2.5 × 10 × 7.5 cm. The inner height of the walls was con-stant (12.5 cm) along the whole length of the staircase. The box wasplaced in a room with constant lighting and isolated from externalnoise. Each mouse was tested individually. The animal was placedon the floor of the staircase with its back to the staircase. The num-ber of stairs climbed and the number of rears were recorded duringa 3-min period. Climbing was defined as each stair on which themouse placed all four paws; rearing was defined as each instancethe mouse rose on hind legs (to sniff the air), either on the stairor against the wall. The number of stairs descended was not takeninto account. Before each test, the animal was removed and the boxcleaned with a diluted alcohol solution to eliminate smells."</li>
<li><strong>Novel object recognition test</strong>: This is a visual recognition memory test that involves measuring the time spent exploring each object.</li>
<li><strong>Y maze test</strong>: This test is used to assess spatial short term memory and involves blocking one arm of the Y-maze in the first trial and then assessing the mouse's memory on subsequent tests. Rationale: "A normal cognitively non-impaired mouse is expected torecognize the old arm as old and spend more time in the new arm."</li>
<li><strong>Rotarod test</strong>: The rotarod tests general motor function and motor learning and measures the time that a mouse can remain walking on a rotating axle without either falling or clenching onto the axle.</li>
</ul><p>So, basically, the investigators injected mice with either saline (negative control), adjuvant only, Gardasil, or Gardasil and pertussis toxin (presumably the positive control), and there were 19(!) animals per experimental group, which is a huge number for most animal studies. The various injections were scaled down from an estimated 40 kg teenaged girl to a 20 g mouse (a 2000-fold difference). The mice received three injections, spaced one day apart. The behavior of the mice, as measured by the parameters above, was then examined at three and six months.</p>
<p>One thing I looked very carefully for in the methods section was something I always look very carefully for in any study of complementary and alternative medicine (CAM). Can you guess what it is? If you're a regular, if you've been reading for a while, I hope that I've imparted enough of my skepticism for you to know right away what I'm talking about. That's right; I looked for any evidence of blinding of observers. I found none. Why is blinding so important? Easy! These measurements are not entirely objective. An observer has to watch the mouse being tested and decide, for instance, what constitutes "immobility" versus "escape-oriented" behaviors in the FST. Without blinding, the observer knows which experimental group each mouse is in, and subtle biases in observation can creep in, without the observer even knowing it. If I had been a reviewer for this paper, I would have immediately noted the lack of any mention of blinding and demanded that the authors clarify whether the observers were blinded or not. If they were not, I would reject the paper.</p>
<p>There's another fatal flaw in the paper as well. Take a look at the statistics section:</p>
<blockquote><p>
Results are expressed as the mean ± SEM. The differences inmean for average immobility time in the FST, the staircase testparameters (number of rearing and stair-climbing events), novelobject recognition and Y maze tests were evaluated by t-test. Significant results were determined as p </p></blockquote>
<p>Those of you unfamiliar with basic statistics (and, believe me, the problem with this passage is very, very basic) won't recognize the problem, but those of you who've taken a basic statistics course will recognize immediately what the problem is here. What is the t-test? No doubt the authors are referring to Student's t-test, which is a test designed to look for differences between two groups. Now, how many groups are being tested? Yes, indeed! It's four. In other words, it's a number of experimental groups for which Student's t-test was never intended. What is the significance of this? Basically, it means that the authors must have compared, pairwise, all the groups. <em>So what?</em> you might ask. Here's the problem. The more comparisons you make, the greater the chance of finding a "statistically significant result" <em>by random chance alone</em>. That's why other statistical tests were developed, specifically the ANOVA test, which would have been the correct test to use to analyze these data. That's another thing I would have insisted on the authors redoing, if I had been a reviewer.</p>
<p>And I would have done it with extreme prejudice, as I have done before in papers I've been asked to review that didn't grasp this <em>very</em> basic bit of statistics.</p>
<p>Now, this might not matter for comparisons for which the calculated p-value is very low, but there are a lot of p-values like 0.02, 0.03, etc., with the cutoff for statistical significance being 0.05. Combine the failure to use an appropriate statistical test with the appropriate post-test to correct for multiple comparisons with the lack of blinding for the behavioral tests, and these p-values are almost certainly not significant. The differences in the FST came with p-values in the range of 0.0001 to 0.025; so some of the differences are probably significant, even if the correct statistical test were used, while others (such as the control versus Gardasil at 6 months, for which p=0.025) are almost certainly not. Regardless, given the apparent lack of blinding of the observers for the behavioral measures, even the differences reported that would likely stand up to an actual correct statistical analysis are very much questionable.</p>
<p>The authors did additional experiments that demonstrated that—surprise! surprise!—Gardasil does induce antibodies to the HPV L1 protein. Amazing. The vaccine works, even in mice! The authors also looked at brain sections of one quarter of the mice in each experimental group, staining brain tissue sections for Iba-1, which is a microglia/macrophage-specific protein that participates in membrane ruffling and phagocytosis in activated microglia; presumably in this case it was being looked at as a measure of inflammation. Of course, if I were reviewing the paper I would have insisted on other measures of inflammation besides staining for just one protein. Basically, they harvested five mice out of each group every month and sectioned their brains. That means a comparison of four groups, each with five mice in them. That's not a particularly robust sample to produce statistically significant results. I'm also a bit cynical about the quantitative claims made for the measurement of Iba-1, given that the selection of "areas of interest" was also manual and apparently also not blinded. </p>
<p>Even given that, the authors found no statistically significant difference between control and Gardasil (p=0.06) but did find a difference between the aluminum adjuvant group and the Gardasil group (p=0.017), which doesn't make a lot of sense if it is the adjuvant that is being blamed for the behavioral changes. Again, this is almost certainly a negative result, given that the authors didn't do the correct statistical test. None of this is to say that there was intentional cooking of the data to give a desired result. However, no observer is entirely objective. That's why blinding of observers is so important in behavioral experiments and experiments that involve human choice of areas to measure on images, even when software is being used to qualify the staining, as studies of immunohistochemistry often do.</p>
<p>Basically, this study is worthless, as it's unblinded and doesn't use the correct statistical analysis. Had I been a reviewer, I would have pointed these issues out and recommended rejecting the paper. I can see why Dr. Poland was probably horrified to discover that this paper was published in his journal. Perhaps he should ask himself how such a travesty could have been published in his journal.</p>
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<span>Tue, 02/16/2016 - 21:00</span>
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Wed, 17 Feb 2016 02:00:46 +0000oracknows22241 at https://scienceblogs.comAfter five years, Bill Maher lets his antivaccine freak flag fly againhttps://scienceblogs.com/insolence/2015/02/09/after-five-years-bill-maher-lets-his-antivaccine-freak-flag-fly-again
<span>After five years, Bill Maher lets his antivaccine freak flag fly again</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p><a href="http://scienceblogs.com/insolence/files/2015/01/Bill_Maher_a_l.jpg"><img src="http://scienceblogs.com/insolence/files/2015/01/Bill_Maher_a_l-450x253.jpg" alt="Bill_Maher_a_l" width="450" height="253" class="aligncenter size-medium wp-image-9266" /></a></p>
<p><strong>Note added 2/10/2015:</strong> <a href="http://scienceblogs.com/insolence/2015/02/10/the-annals-of-im-not-anti-vaccine-part-13-nobody-wants-to-admit-to-being-antivaccine/">I've posted a followup in response to the skeptics who defend Bill Maher</a>.</p>
<p>A couple of weeks ago, I noted the <a href="http://scienceblogs.com/insolence/2015/01/19/bill-maher-still-an-antivaccine-wingnut-after-all-these-years/">return of the antivaccine wingnut side of Bill Maher</a>, after a (relative) absence of several years, dating back, most likely, to the thorough spanking he endured for spouting off his antivaccine pseudoscience during the H1N1 pandemic. This well-deserved mockery included Bob Costas taunting him on his own show with a sarcastic, "<a href="http://scienceblogs.com/insolence/2008/02/24/is-bill-maher-really-that-ignorant-part-3/">Oh, come on, Superman!</a>" in response to his apparent belief that diet and lifestyle alone would protect him from the flu, as well as Chris Matthews doing the same thing, <a href="http://scienceblogs.com/insolence/2015/01/19/bill-maher-still-an-antivaccine-wingnut-after-all-these-years/">likening Bill Maher to a celebrity Scientologist</a> denying psychiatry to his face. Then <a href="http://scienceblogs.com/insolence/2009/10/16/from-one-skeptic-to-the-2009-recipient-o/">Michael Shermer took him on</a>, gently remonstrating with him, which led Maher to <a href="http://scienceblogs.com/insolence/2009/11/17/bill-maher-flames-out-over-vaccines/">go full mental jacket trying to defend himself</a>. He was even <a href="http://scienceblogs.com/insolence/2009/10/12/the-2009-recipient-of-the-richard-dawkin/">slapped down by Senator Bill Frist</a> for saying he doesn't believe in vaccines or "Western medicine." Of course, given that I've been <a href="http://oracknows.blogspot.com/2005/12/bill-maher-anti-vax-wingnut.html">covering Maher's antivaccine proclivities</a> for a <a href="http://oracknows.blogspot.com/2005/03/is-bill-maher-really-that-ignorant_07.html">decade now</a>, I was under no illusion that he had suddenly gone a conversion to science. Rather, I just thought (<a href="http://scienceblogs.com/insolence/2015/01/19/bill-maher-still-an-antivaccine-wingnut-after-all-these-years/">correctly</a>, as it turns out) that he was laying low, licking his wounds. So when he went anti-flu vaccine a couple of weeks ago, I wondered if that was a harbinger of things to come.</p>
<p>Then, earlier this week, I saw an editorial by Andrew Kirell, <a href="http://www.mediaite.com/tv/will-bill-maher-address-his-long-history-of-vaccine-skepticism-this-week/">Will Bill Maher Address His Long History of Vaccine Skepticism This Week?</a> Kirell concluded his op-ed asking:</p>
<blockquote><p>
And that brings us to this week. His Real Time panel includes no doctors, but features two conservative pundits and a journalist — any of whom will likely take the opportunity to prod Maher in light of this week’s news.</p>
<p>So will Maher address his history on the matter and say something controversial? It seems unavoidable.
</p></blockquote>
<p>If the episode two weeks ago was just the <em>hors d'oeuvre</em>, this week's episode of <a href="http://www.hbo.com/real-time-with-bill-maher#/">Real Time With Bill Maher</a> was the main course of full-on antivaccine wingnuttery. Seriously, this might well be the worst Maher's ever been with respect to science, yoking in appeals to ignorance, specious comparisons with anthropogenic global warming, various anti-pharma rants, and, of course, GMO hysteria. Here's <a href="http://youtu.be/B7yvI0tu3Ho">the offending segment</a> (although Maher did mention earlier in the show that he's not "antivaccine" just "anti-flu vaccine"):</p>
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<!--more--><p>For advocates of science, this is painful to watch, as Maher and his guests rubbish vaccines, "Western" medicine, GMOs, big pharma, Monsanto, and all the usual suspects that cranks and quacks attack. Before I address the specific misinformation and pseudoscience promoted in this episode, let me first note that clearly Maher must have learned something from previous embarrassments. For example, his exploratory rant against this year's flu vaccine (whose efficacy is, unfortunately, less than usual and disappointing) was <a href="http://scienceblogs.com/insolence/2015/01/19/bill-maher-still-an-antivaccine-wingnut-after-all-these-years/">easily countered by Atul Gawande</a>, a real physician and researcher, just as Bill Frist, a real physician, <a href="http://scienceblogs.com/insolence/2009/10/12/the-2009-recipient-of-the-richard-dawkin/">countered him before</a>. Heck, even <a href="http://scienceblogs.com/insolence/2008/02/24/is-bill-maher-really-that-ignorant-part-3/">Bob Costas</a> and <a href="http://scienceblogs.com/insolence/2009/10/17/the-2009-recipient-of-the-richard-dawkin-1/">Chris Matthews</a> were able to counter Maher's misinformation. This time around, Maher clearly made sure there was no one who was likely to contradict his quackery-laden views or take him to task for spreading antivaccine pseudoscience on his show.</p>
<p>First up, there was Marianne Williamson, who <a href="http://www.marianneforcongress.com">apparently ran for Congress last year</a>. But there's more than that. I had never heard of her before, but apparently she's some sort of <a href="http://marianne.com">author and "spiritual teacher."</a> Her <a href="http://blog.marianne.com/journal/index.php">blog</a> is New-Agey and woo-ey, <a href="http://www.bloomberg.com/bw/articles/2014-04-10/marianne-williamson-californias-new-age-contender-for-congress">as is she</a>, as her <a href="https://www.facebook.com/williamsonmarianne/info?tab=page_info">Facebook profile</a> shows:</p>
<blockquote><p>
Marianne Williamson is an internationally acclaimed spiritual teacher. Her first book, A Return to Love: Reflections on the Principles of A COURSE IN MIRACLES, is considered a must-read of The New Spirituality. A paragraph from that book, beginning "Our deepest fear is not that we are inadequate. Our deepest fear is that we are powerful beyond measure" - often misattributed to Nelson Mandela's Inaugural address - is considered an anthem for a contemporary generation of seekers.
</p></blockquote>
<p>If you don't believe me, then read this interview with her about the "<a href="http://consciouslifenews.com/law-divine-compensation-marianne-williamson-book-excerpt/">Law of Divine Compensation</a>."</p>
<p>More tellingly, apparently before her appearance on Maher's show she had been posting links to articles about the Gates Foundation. Or something. Whatever the reason, on February 1 felt the need to <a href="https://www.facebook.com/williamsonmarianne/posts/169170463129485">post this</a>, where she states that she took down several posts, apparently about vaccines, because her fans were trashing her. At least, that's all I could figure out from the comments:</p>
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<div class="fb-xfbml-parse-ignore"><a href="https://www.facebook.com/williamsonmarianne/posts/169170463129485">Post</a> by <a href="https://www.facebook.com/williamsonmarianne">Marianne Williamson</a>.</div>
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<p>Many of the comments after are a veritable hive of antivaccine sentiment, complete with links to articles by antivaccine loons like Gary Null, Sherry Tenpenny, and Mike Adams. If Williamson attracts such an antivaccine crowd, one has to wonder, particularly in light of her performance on Maher's show. Certainly, even if she is not antivaccine, she is too clueless or doesn't care enough to make a defense of vaccination.</p>
<p>Another of Maher's guests is <a href="http://www.theblaze.com/author/amy-holmes/">Amy Holmes</a> of <a href="http://www.theblaze.com">The Blaze</a>, which Glenn Beck's TV channel. Obviously, that's a <a href="http://www.mediaite.com/online/glenn-becks-gbtv-names-cnns-amy-holmes-news-anchor-for-the-blaze/">bad sign right there</a>, given Glenn Beck's propensity for conspiracy mongering. I couldn't find any evidence that she's ever voiced antivaccine views before (or, for that matter, anything much at all about vaccines). So we have another reporter, this time working for <a href="http://www.theblaze.com/shows/the-glenn-beck-program/">Glenn Beck</a>. This is not a good indication that she has any scientific background.</p>
<p>Finally, there is conservative columnist <a href="http://www.weeklystandard.com/author/john-mccormack">John McCormack</a> of the <a href="http://www.weeklystandard.com">Weekly Standard</a>. Contrary to a couple of conservatives who voiced some antivaccine-sympathetic nonsense last week, McCormack is the only one on Maher's panel who showed a modicum of sense, although he was not willing to challenge Maher that strongly, and one of his challenges was a politically motivated misfire expressing anthropogenic global climate change denialism, as you will see. It's basically fighting pseudoscience with pseudoscience, and that doesn't really make a particularly good case.</p>
<p>You know things are not going to go well, scientifically speaking, when, right off the bat, Maher introduces the segment by referring to the meeasles outbreak as the "topic that's getting everybody crazy in America" and then saying:</p>
<blockquote><p>
When I start these conversations, I always have to say: I'm not an antivaxer. I never have been. I'm an anti-flu shot guy I think that's bullshit, and the fact that it was only 23% effective this week bears that out. But if Ebola was airborne, I'd get the vaccine tomorrow.
</p></blockquote>
<p><a href="http://scienceblogs.com/insolence/2015/01/16/cdc-did-not-admit-flu-vacine-does-not-work/">Wrong, wrong, wrong, wrong</a>. Good thing for Maher that Atul Gawande wasn't there to school him about the flu vaccine as <a href="http://scienceblogs.com/insolence/2015/01/19/bill-maher-still-an-antivaccine-wingnut-after-all-these-years/">he did last time</a> as Maher deserved to be schooled. Of course, Gawande is too nice to school Maher as he needs to be schooled and Maher would never allow anyone with both the knowledge and the necessary willingness to call Maher out properly to carry out the task on his show. It is, after all, his show. In any case, I've documented more times than I'd care to count that Maher is indeed antivaccine to the core—and pro-quackery to the core. Maher labors under the delusion that he is more rational than everybody else, and his smugness and condescension drip from his very essence, oozing from the television (or computer screen, depending on what you're watching him on).</p>
<p>It gets even worse when Maher immediately starts complaining about the "attitude of the media," which he characterized as "just a lot of shut the fuck up." He even compared it to the first weeks of the Iraq war. This lead Williamson to chime in that the implication was that "if you had any skepticism whatsoever, you were antiscience." Of course, Bill Maher is anti-science with respect to vaccines, even though he views himself as totally pro-science. So he lapped this up, particularly when she followed it up with the self-serving Maher-approved observation that there is a "difference between having skepticism about science and having skepticism about the pharmaceutical industry." Truly her stupid did burn brightly. It burns brighter still. Even as she touts that she vaccinated her children, she goes on about how the government has "earned our distrust" and how the "government has suppressed information" and medicine has done the same, she bristles at being called antiscience for being suspicious of the pharmaceutical industry. Her conclusion? She says that the answer is "not to call us kooks" but for the government and pharmaceutical industry to "get their acts together."</p>
<p>Of course, this is a tactic taken straight from the playbook of the antivaccine movement, to conflate (disingenuously) reasonable suspicion of the pharmaceutical industry's previous misdeeds with suspicions of vaccines. They are not the same thing, nor is one as reasonable as the other. Whatever misdeeds the pharmaceutical industry might be guilty of, they do not cast doubt on the safety and efficacy of vaccines. There is plenty of independent evidence to support the conclusions that vaccines do not cause autism, they do not cause neurodevelopmental disorders, and they do not cause sudden infant death syndrome, allergic conditions, or any of the other problems frequently ascribed to them by antivaccinationists. No matter how much the government or the pharmaceutical industry "gets its act together" it's never, ever enough for kooks like Marianne Williamson. (I couldn't resist.) Also, the claim that you "can't question" is a favorite cry of the crank.</p>
<p><a href="http://youtu.be/l8ukak8P2vY">Help, help, I'm being repressed</a>!</p>
<p>Unfortunately, Amy Holmes can't resist adding to the stupid of the whole affair. She characterizes the news coverage as "gotcha politics," in which Governor Chris Christie and Senator Rand Paul are made to look like kooks or "anti-science" (Holmes even does air scare quotes to emphasize the point), a comparison that literally made me do the rare double facepalm upon hearing it and practically shouting at the television. No wonder this woman works for Glenn Beck! She then points out that 48 states allow parents to have religious and/or personal belief exemptions. Yes, that's true, but so what? It's bad policy, and 48 states have bad policy. In any case, she tries to burnish her science bona fides by saying that she "doesn't worship at the church of Jenny McCarthy" as she describes the case of a woman with a child with leukemia, but her overall attitude is that it's "gotcha politics" to have called out Gov. Christie and Sen. Paul for their antivaccine nonsense.</p>
<p>It isn't, and it isn't "gotcha politics" to call Sen. Rand Paul antivaccine. <a href="http://scienceblogs.com/insolence/2015/02/03/is-republican-party-becoming-antivaccine-party/">He is</a>.</p>
<p>At this point, John McCormack dives in as the seeming voice of reason, which is good. Unfortunately, he couldn't resist making the claim that this is not a Republican problem but more of a "liberal problem." It's not. Antivaccinationism is very at home <a href="http://scienceblogs.com/insolence/2013/12/18/why-are-antivaccinationists-so-at-home-with-libertarianism/">among libertarians</a> and <a href="http://scienceblogs.com/insolence/2012/07/16/battling-antivaccinationists-at-freedomfest/">conservatives</a>, and there's no evidence that this is a "liberal problem," the stereotype notwithstanding. As I've said so many times before, antivaccinationism is the quackery and pseudoscience that transcends political boundaries. By trying to paint antivaccine beliefs as more a "liberal" problem, McCormack shows his true agenda. (Hint: It's not to defend science.)</p>
<p>If you want more evidence of this, then check out the next exchange. First, Maher makes this ludicrous analogy:</p>
<blockquote><p>
The analogy that I see all the time is that if you ask any questions, you are the same thing as a global warming denier. I think this is a very bad analogy, because I don’t think all science is alike. I think climate science is rather straightforward because you’re dealing with the earth. It’s a rock. I'm not saying I know how to deal with it, but climate scientists, from the very beginning, have pretty much said the same thing, and their predictions have pretty much come true. It’s atmospherics, and it’s geology, and chemistry. That’s not true of the medical industry. I mean, they’ve had to retract a million things because the human body is infinitely more mysterious. People get cancer, and doctors just don't know why. They just don't know why, and they don't know how to fix it. And they put mercury in my teeth. My father had ulcers and they treated it wrong when I was a kid. Thalidomide. I mean I could go on about how many times they have been wrong. To compare those two science is, I think, just wrong.
</p></blockquote>
<p>And <a href="http://knowyourmeme.com/memes/fucking-magnets-how-do-they-work">magnets, how do they work?</a></p>
<p>Seriously. This is nothing more than the "science was wrong before" gambit. Let's just put it this way. Physics has gone through many iterations and has had to "admit" that many of its prior theories were wrong. Does Maher doubt, for instance, the theory of relativity, which supplanted Newtonian physics? His analogy is just so utterly, breathakingly stupid that I did the double double facepalm upon hearing it. In fact, doubting the safety and efficacy of vaccines is <em>very much</em> like climate science denialism. Both are areas of science that are well accepted by the scientific community and backed by enormous quantities of evidence.</p>
<p>Here's where McCormack goes off the rails. He mentions that there is an M.I.T. professor, Richard Lindzen, who's a climate skeptic, but there are no such professors that are vaccine skeptics. Of course, being a professor doesn't mean you're not a denialist, and in fact <a href="http://www.realclimate.org/index.php/archives/2012/03/misrepresentation-from-lindzen/">Lindzen is a denialist</a>. He's also the <a href="http://www.realclimate.org/index.php/archives/2007/04/lindzen-in-newsweek/">beneficiary of oil industry money</a>, which is amusing because it led Maher to say the one thing he said in this entire segment that is mostly correct, namely that most climate "skeptics" have ties to industry. McCormack is also just plain wrong that there are no professors who are "vaccine skeptics." For instance, there's Christopher Shaw, who is a Professor of Ophthalmology and Visual Sciences at the University of British Columbia whose research specializes in neurodegenerative diseases. He's also an antivaccine crank who thinks aluminum adjuvants in vaccines <a href="http://www.neuroscience.ubc.ca/shaw.htm">cause Alzheimer's disease and autism</a>. He's appeared in the antivaccine propaganda movie <a href="http://scienceblogs.com/insolence/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">The Greater Good</a> and has been interjected himself into a sad case of an 18-year-old female who died suddenly, <a href="http://scienceblogs.com/insolence/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">trying to blame it on Gardasil</a>. Oh, yes, Mr. McCormack. There are definitely antivaccine professors out there. They are very much like AGW denialist professors in their <em>modus operandi</em>.</p>
<p>Maher's next argument is just plain dumb. He decides he's going to liken vaccines to antibiotics and ask, "Can you just do too much of a good thing?" and "Is it limitless? Is there no amount that is too much?" At another point, he seems to imply that scientists were surprised that antibiotic resistance has become so widespread, when in fact it was scientist warning about overuse of antibiotics who foresaw this problem. This leads Williamson to repeat the tired old antivaccine trope of "too many, too soon" in the form of JAQing off. Maher feeds off of that by acknowledging that vaccines don't cause autism and that he "accepts that," but then pivots to the <a href="http://scienceblogs.com/insolence/2009/08/24/its-so-cute-when-anti-vaxers-try-to/">classic antivaccine trope</a> that there are no long term studies of vaccinated versus unvaccinated children and "wonders" if people who've had a lot of vaccine have "robust immune systems." He links this to more diagnoses of allergies, autoimmune diseases, and the like, in a classic bit of JAQing off in which he says he isn't claiming that vaccines are responsible for this. He's just asking questions, you know—and confusing correlation with causation.</p>
<p>As my good bud <a href="http://scienceblogs.com/denialism/2015/02/07/bill-maher-is-an-astonishingly-anti-science-anti-vax-crank/">Mark Hoofnagle notes</a>, he even does some serious mental gymnastics in which he goes on about how he thinks that if you don't use your immune system, you'll lose it. The problem, of course, is that vaccines activate the immune system by stimulating it with the same antigens that one finds in the pathogens that cause disease. They wouldn't work if that weren't what they do. So Maher can't even keep a coherent train of thought. On the one hand, supposedly we have all these autoimmune diseases, presumably because vaccines stimulate the immune system too much, but then people who have been vaccinated don't have as "robust an immune system." Which is it Bill? And do you have the slightest clue how stupid about medicine you sound?</p>
<p>Obviously not.</p>
<p>At this point I can't resist a little dig at Amy Holmes' ignorance about smallpox. She notes that she has had a smallpox vaccine because she's was born out of the country and notes ("thank goodness") that we are "eradicating smallpox." News flash for Ms. Holmes: We are not eradicating smallpox. We eradicated it decades ago, thanks to vaccines. There have been no natural cases since 1977, and the last known case was due to a laboratory accident in 1978. It's <a href="http://www.who.int/csr/disease/smallpox/en/">been 37 years since there's been a recorded case of smallpox</a>, because of vaccines. It gets even worse. Maher makes an incoherent analogy to testosterone supplementation, in which such supplementation "makes your balls shrink." He then analogizes that to vaccines and the immune system, implying that if you use vaccines your immune system thinks it doesn't have to work so hard. Again, does this clown even know how vaccines work?</p>
<p>Maher also complains that he's never had a "Western doctor" ask him about his diet. Really? If his anecdote is to be believed, then let me point out my anecdote. Every doctor I've ever had asked me about my diet. I also note that, until the last several years, I was actually pretty thin. Twenty years ago, I was actually skinny. However, as I got into my 40s and hit 50, biology betrayed me (as it is wont to do as one gets older) and, although I'm not fat, I'm no longer thin. Around that time, when I went from being thin to being average to being slightly overweight, lo and behold! My doctor started asking me about diet and lifestyle. </p>
<p>This leads to a curious rant about GMOs and an attack on Monsanto, or, as I like to call it, argumentum ad Monsanto. At this point, McCormack argues that GMOs have been a great force for reducing world hunger, which is undoubtedly true. Maher dismisses such arguments with a jaunty, "But I'm not a starving child in Africa. If I were a starving child, then, yes, I'd eat a GMO." McCormack then asks what studies show that GMOs are harmful, which leads Williamson and Maher to become condescendingly dismissive, with "WTF? Are you kidding me?" looks on their faces. Of course, as I've described before, the only studies that have claimed to show dangers from GMOs are studies <a href="http://scienceblogs.com/insolence/2012/09/24/bad-science-on-gmos-it-reminds-me-of-the-antivaccine-movement/">done by anti-GMO advocates</a> and studies with <a href="http://scienceblogs.com/insolence/2013/06/17/bad-science-about-gmos-it-reminds-me-of-the-antivaccine-movement-revisited/">very poor design</a>. These are the sorts of studies that evidently impress Maher and Williamson, utter crap.</p>
<p>Maher believes himself to be the real pro-science advocate. He is about as wrong as wrong can be. He is anti-vaccine, anti-"Western medicine," and in general antiscience, except for a limited number of areas of science that fit in with his ideological biases. As such, he's an object lesson in how one can be intelligent and anti-science at the same time. He's also an object lesson in how being an atheist and being pro-science are related only by coincidence. I had thought that Maher might have been sufficiently chastened by the spanking he received in 2009 and 2010 about his antivaccine stylings. Apparently five years have been enough time for his antivaccine freak flag to fly again.</p>
<p>He is no skeptic. He is no pro-science advocate. He's an occasionally funny political comedian with delusions of grandeur with respect to his own rationality.</p>
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Mon, 09 Feb 2015 00:00:54 +0000oracknows21984 at https://scienceblogs.comAnd now death by Gardasil? Again, not so fast...https://scienceblogs.com/insolence/2012/10/31/and-now-death-by-gardasil-again-not-so-fast
<span>And now death by Gardasil? Again, not so fast...</span>
<div class="field field--name-body field--type-text-with-summary field--label-hidden field--item"><p>I guess this is in effect part two of <a href="http://scienceblogs.com/insolence/2012/10/30/ovarian-failure-caused-by-gardasil-not-so-fast/">yesterday's post</a>. Regular daily readers (and you are a regular daily reader, aren't you?) will remember that yesterday I commented on the recent uptick in anti-Gardasil vaccine rhetoric coming from the antivaccine crank blog Age of Autism and other sources, in the process deconstructing speculation masquerading as a case report allegedly indicting the quadrivalent HPV vaccine as a potential cause of premature ovarian failure in a 16 year old Australian girl. The article was so bad and so biased that I couldn't believe BMJ Case Reports published it in the form it did. It's almost enough to make me take back all those <a href="http://scienceblogs.com/insolence/2011/01/06/piltdown-medicine-andrew-wakefields-scie/">nice things I said about the BMJ</a> in the wake of its publication of Brian Deer's expose of Andrew Wakefield's fraud, but in reality the publication of one bad piece of antivaccine propaganda doesn't invalidate all the good the BMJ did by commissioning Brian Deer to lay the results of his investigation on the line.</p>
<p>You'll also recall that I noted another article that's popping up on antivaccine websites. Not content with trying to scare parents and girls with stories of Gardasil robbing young women of their womanhood, antivaccinationists now want to scare parents with stories of Gardasil robbing girls of their lives. So it was that the antivaccine propaganda blog <a href="http://www.ageofautism.com/2012/10/age-of-autism-science-summary-death-after-quadrivalent-human-papillomavirus-hpv-vaccination.html" rel="nofollow">touting an article</a> by the latest scientists to have drunk the pseudoscience Kool Aid that is antivax, Lucija Tomljenovic and Christopher A. Shaw. AoA was joined by the rabidly anti-Gardasil antivaccine group <a href="http://sanevax.org/death-after-quadrivalent-human-papillomavirus-hpv-vaccine-causal-or-coincidental/" rel="nofollow">(IN)SANE Vax, Inc.</a>, the <a href="http://www.greatergoodmovie.org/news-views/1596/" rel="nofollow">blog</a> for the antivaccine movie <a href="http://scienceblogs.com/insolence/2011/11/18/anti-vaccine-propaganda-lands-in-new-yor/">The Greater Good</a>, <a href="http://gaia-health.com/gaia-blog/2012-10-25/gardasil-is-probable-cause-of-girls-deaths-brain-histology-study/" rel="nofollow">Gaia Health</a>, and others. That's no surprise, because we've met Dr. Shaw before. He was a key player in that particular bit of propaganda, likening vaccines to part of a toxic soup of chemicals that contributes to autism. We've also met Tomljenovic before. Along with Shaw as her co-author, she wrote one of the silliest attempts at blaming vaccines for autism that I've seen in a long time (and I've seen many, many very silly attempts to blame vaccines for autism over the years). Basically, Tomljenovic and Shaw tried to "prove" that the rising autism prevalence is due to the use of aluminum adjuvants in vaccines and failed utterly, <a href="http://scienceblogs.com/insolence/2011/12/08/and-global-warming-is-caused-by-the-decr/">hilariously confusing correlation with causation</a> in a textbook example of how not to draw inferences from statistical data.</p>
<!--more--><p>Finally, we've also <a href="http://scienceblogs.com/insolence/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">met Shaw before</a> when he testified before an investigative committee looking into the death of an 18-year-old woman named Jasmine Renata in New Zealand. Basically, Jasmine's mother was convinced that her daughter had died because of the HPV vaccine rather than a much more plausible and likely cause, an undiagnosed heart conduction defect. He claimed that he found HPV DNA associated with aluminum in Jasmine's brain, which would be a real pharmacokinetic and stoichiometric feat if true given the tiny amount of HPV DNA found in the HPV vaccine. It's such a small amount that it takes a remarkably sensitive (and possibly nonspecific) <a href="http://scienceblogs.com/insolence/2011/09/06/oh-no-theres-dna-in-my-gardasil/">nested quantitative real time PCR technique</a> promoted by a <a href="http://scienceblogs.com/insolence/2011/10/12/it-couldnt-happen-to-a-nicer-guy-1/">discredited pathologist</a> named Sin Hang Lee working with (IN)SANE Vax to detect it. So annoyed was Dr. Shaw at my criticism of his testimony that he even <a href="http://scienceblogs.com/insolence/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">briefly showed up in the comments of this post</a>.</p>
<p>All of this brings us to the latest spew by Tomljenovic and Shaw, which was published in a journal I had never heard of, <a href="http://www.omicsgroup.org/journals/proahome.php">Pharmaceutical Regulatory Affairs</a>, and entitled <a href="http://sanevax.org/wp-content/uploads/2012/10/Tomljenovic-Shaw-Gardasil-Causal-Coincidental-2167-7689-S12-001.pdf" rel="nofollow">Death after quadrivalent human papillomavirus (HPV) vaccination: Causal or coincidental?</a> Guess which side Tomljenovic and Shaw want to persuade you to accept as the correct side of this question? (Hint: It ain't the science-based side.)</p>
<p>Basically, this article consists of two case reports. Case 1 is very clearly Jasmine Renata, <a href="http://scienceblogs.com/insolence/2012/08/09/a-sad-premature-death-cynically-used-by-antivaccinationists-to-attack-gardasil/">whose case I discussed in depth</a> about three months ago. The case description matches almost exactly:</p>
<blockquote><p>A 19-year-old female without a relevant medical history and taking no drugs expired in her sleep, approximately 6 months after her third and final qHPV vaccine booster and following exacerbation of initial vaccination-related symptoms. She had last been seen alive by her parents the previous evening. Her symptoms started after the first qHPV injection when she developed warts on her hand that persisted throughout the vaccination period. In addition, she suffered from unexplained fatigue, muscle weakness, tachycardia, chest pain, tingling in extremities, irritability, mental confusion and periods of amnesia (memory lapses). The autopsy was unremarkable and failed to determine the exact cause of death. Internal examination revealed some minor changes involving the gallbladder and the uterine cervix (both of which on further examination by microbiological studies and histology revealed no significant disease). After a full autopsy no major abnormality was found anatomically, microbiologically or toxicologically that might have been regarded as a potential cause of death. Histological analysis of the brain hippocampus, cerebellum and watershed cortex allegedly revealed no evidence of neuronal loss or neuroinflammatory changes. However, the autopsy report did not specify which immune antibodies and stains were used for histological investigations.</p></blockquote>
<p>Case 2 is very almost certainly another <em>cause célèbre</em> in the antivaccine movement, Annabelle Morin of Quebec, who died in 2008 and whose parents are <a href="http://www.lifesitenews.com/news/parents-sue-after-quebec-teen-dies-following-gardasil-vaccination" rel="nofollow">suing Merck for her death</a> after apparently encountered <a href="http://articles.mercola.com/sites/articles/archive/2011/11/29/hpv-vaccine-risks.aspx?e_cid=20111129_DNL_art_1" rel="nofollow">Joe Mercola's fear mongering about Gardasil</a> online. The clinical history presented sounds very much like Morin's:</p>
<blockquote><p>A 14-year-old female with a previous history of migraines and oral contraceptive use developed more severe migraines, speech problems, dizziness, weakness, inability to walk, depressed consciousness, confusion, amnesia and vomiting 14 days after receiving her first qHPV vaccine injection. These symptoms gradually resolved. However, 15 days after her second qHPV vaccine booster she was found unconscious in her bathtub by her mother 30 minutes after she had entered the bathroom to have a shower. Emergency help was summoned and arrived quickly. Resuscitation efforts were attempted. The paramedic noted that the patient was found without a pulse. Upon arrival at the hospital and approximately 30 minutes later, the patient suffered cardiac arrest. Resuscitation was terminated approximately 40 minutes later and the patient was pronounced dead.</p></blockquote>
<p>Given the resemblance, it must be Annabelle they're writing about here.</p>
<p>The first thing I noticed here s that Shaw's story seems to be...evolving. What do I mean? Simple. Go back and look at what he said about Jasmine originally. He claimed that he <a href="http://www.stuff.co.nz/dominion-post/news/7445193/Foreign-DNA-found-in-teenagers-blood">found HPV DNA</a> in her blood and spleen after her death, saying to the inquest into Jasmine Renata's death:</p>
<blockquote><p>He said it was not the result of a natural HPV infection, most likely the DNA was bound to aluminium which was also found in Jasmine.</p>
<p>“The HPV gene is foreign DNA and its detection six months after injection is not normal,’’ he told the inquest.</p>
<p>He said the DNA may cause a reaction that could lead to lethal shock although it was not known if it caused her death but it needed further investigation.</p>
<p>He said it was not known if it was the cause of death but it needed further investigation.</p></blockquote>
<p>So let's see. Originally, Shaw was saying that somehow the aluminum adjuvant in the HPV vaccine somehow complexed with HPV DNA from the vaccine in order to work its evil effects. He said this even though, as I explained above, the amount of HPV DNA in a dose of Gardasil is so small that it takes ridiculously sensitive PCR techniques to detect it, and, even then, I'm not entirely convinced that what Lee found really was HPV DNA and not a contaminant. We're probably talking nanogram, if not picogram, quantities of DNA. It defies basic chemistry and plausibility to propose that such a minuscule amount of DNA not packaged in a virus or other delivery vector could cause such a reaction. It makes one wonder, it does. Did Shaw realize that his original story didn't pass the smell test? Did he realize that molecular biologists were rolling in the floor with laughter or snorting at him with contempt over this explanation? Did he actually take my post to heart? Think of it. His manuscript is listed as having originally been submitted on September 13, 2012 and accepted for publication on October 2, with a publication date of October 4. While noting that that's a mighty fast turnaround time (which makes me wonder about the quality of the peer review for this journal), I also note that it's also over a month after my post. Sure, I could have an overinflated sense of my own importance. It wouldn't be the first time I've been accused of that. But, still, I wonder.</p>
<p>Whatever the reasons, Tomljenovic and Shaw are now claiming something somewhat different but only marginally more plausible. I'm not a neuroscientist, nor am I a pathologist, but one thing I can't help but note is that neither Tomljenovic and Shaw are pathologists, either. In particular, neither of them are neuropathologists. If there's one thing I know about neuropathology, it's that it's tricky. It's not trivial to get antibodies to work properly on brain tissue, nor is it a trivial matter to interpret. None of this seems to have induced our not-so-dynamic duo to have recruited a neuropathologist to assist them with interpretation of the immunohistochemical stains of the brain sections. Were I a reviewer for this paper, I would have recommended not publishing it without a neuropathologist as an author. None of this keeps the authors from boldly proclaiming that neuroinflammation caused by the HPV-16L1 antigen is strongly implicated as the cause of these girls' deaths through an autoimmune vasculitis due to the deposition of HPV-16L1</p>
<p>Not so fast, there pardner.</p>
<p>The first thing to consider is the biological plausibility of the HPV-16L1 protein somehow depositing in the brain vasculature in order to be able to cause an immune reaction. Each dose of Gardasil contains approximately <a href="http://www.rxlist.com/gardasil-drug.htm">20 μg HPV-16L1 protein</a>, which is injected locally intramuscularly. While this is clearly not difficult to detect the way the trace DNA left over in Gardasil is, it's still by no means a lot of protein. Yet, according to our not-so-dynamic duo, this tiny amount of protein caused this:</p>
<blockquote><p>
In both cases, the autopsy revealed no anatomical, microbiological nor toxicological findings that might have explained the death of the individuals. In contrast, our IHC analysis showed evidence of an autoimmune vasculitis potentially triggered by the cross-reactive HPV-16L1 antibodies binding to the wall of cerebral blood vessels in all examined brain samples. We also detected the presence of HPV-16L1 particles within the cerebral vasculature with some HPV-16L1 particles adhering to the blood vessel walls. HPV-18L1 antibodies did not bind to cerebral blood vessels nor any other neural tissues. IHC also showed increased T-cell signalling and marked activation of the classical antibody-dependent complement pathway in cerebral vascular tissues from both cases. This pattern of complement activation in the absence of an active brain infection indicates an abnormal triggering of the immune response in which the immune attack is directed towards self-tissue.
</p></blockquote>
<p>Here's the thing. The autopsy didn't show any abnormalities in Jasmine's brain that could account for her death. The same was true of Annabelle. If these girls had an immune-based vasculitis, it should have been visible as severe inflammation in the brain tissues on normal H&E sections (sections stained with the usual blue and pink dye and no special immunohistochemical stains). Then, the IHC would reveal the potential cause. For instance, in Case 1, Tomljenovic and Shaw note that the pathologist found no signs of neuronal loss or neuroinflammatory changes but that it wasn't reported which antibodies were used. More than likely the answer is none; routine sections would just be H&E, with IHC reserved for cases in which an abnormality was noted on H&E. In Case 2, Tomljenovic and Shaw bemoaned how the pathologist didn't use any specific antibodies for inflammatory markers but rather noted changes consistent with terminal ischemic-hypoxic encephalopathy. What that means is that the pathologist thought that the changes he observed were consistent with the brain's having suffered a significant period of time with low blood flow, rendering it ischemic, something that could have happened if the girl had a very low blood pressure for a while before she died.</p>
<p>So what we have is a non-pathologist, who clearly doesn't know how to interpret common pathological findings, throwing every inflammation-related antibody in the book at the brain section and then concluding that there is some sort of neuroinflammation because he sees staining in the blood vessels in the brain. There's one huge problem, though, and Tomljenovic and Shaw basically admit it:</p>
<blockquote><p>
The obvious limitations of our study are that the tissues examined represent two individuals against which there were no control samples. For this reason, we could not obtain a quantitative measure of immunoreactivity. We aim in the future to further corroborate our findings by examining brain tissues from other cases of sudden and unexplained death following HPV vaccination, as well as control brain tissue from age-matched individuals who clearly died from nonvaccination related causes. Nonetheless, the marked resemblance in immunostaining patterns for all immunohistological markers in brain tissue specimens in the present two cases (i.e., compare Figures 1-4), as well as the similarity between their symptoms and those noted on VAERS reports related to post-HPV vaccination vasculopathies (some of which were medically ascertained cases; Tables 2 and 3), strongly support our present conclusions.
</p></blockquote>
<p>Uh, no they don't. Tomljenovic and Shaw's findings could just as well be nonspecific immunostaining. Not only didn't they examine sections of age-matched normal brains as controls (the very minimum they could have done), but, as far as I can tell from this paper, they neglected to do some <a href="http://www.rndsystems.com/literature_ihc_icc_controls.aspx">very basic controls for any IHC experiment</a>. Specifically, I don't see any reference to a "no primary antibody" control or an isotype control. In the absence of such controls, there is no compelling evidence that the staining that Tomljenovic and Shaw are reporting is anything other than due to nonspecific interactions between the antibody and cellular structures. This is a lot more common than one might think. Many are the physicians and scientists who have tried to do IHC for a research project and found that it's nowhere near as cookbook as the antibody and reagent manufacturers would like you to think from their protocols. It's one reason why IHC is often referred to as an art as much as a science. it often takes a lot of trial and error to get it right, even with commercial antibodies with seemingly well-defined protocols from the manufacturer.</p>
<p>Another thing they could have done to convince skeptics like me is something called an adsorption control Basically, that means incubating the primary antibody with the antigen it's supposed to detect. If the interaction being observed is specific, then adsorbing it this way should eliminate the staining, because the specific antibody will complex with the antigen. Finally, there should be a positive control. For HPV proteins, this would mean perhaps cervical cancer lesions known to be due to HPV infection. I will admit that a lot of investigators don't report all of these controls, but they usually report at least a couple of them, usually the isotype control and the no primary antibody control. In particular, for a novel or unusual finding (such as the finding of HPV-16L1 in the neurovasculature), more controls need to be done. Also, reviewers will give a skilled pathologist a bit more of the benefit of the doubt if he doesn't present all controls, because, well, he's a pathologist and does this sort of thing every day for a living in real patient samples. Tomljenovic and Shaw don't rate that sort of benefit of the doubt. They should show their work, but do not. If I had to guess, what we're seing in their IHC sections is nonspecific staining.</p>
<p>That having been said, I'm not a pathologist, either, although I've done a fair amount of IHC in my laboratory. The difference, of course, is that I'm not diagnosing patients. In any case, as Dirty Harry once said, "A man's got to know his limitations." I know mine. I know that I could be wrong. So I'd love it if any of my readers are pathologists, particularly neuropathologists, could comment. In particular, the choice of antibodies by Tomljenovic and Shaw seems to include antibodies that are mainly used for research and not routinely used in clinical specimens for actual diagnosis, which means even more that, even if what we're seeing is not nonspecific, its significance is unclear. Still, given the tiny amount of HPV-16L1 in each Gardasil dose, I'd bet that the finding of this protein in the neurovasculature of these girls is almost certainly nonspecific staining.</p>
<p>Basically, Tomljenovic and Shaw have become the latest not-so-dynamic antivaccine duo trying to lend scientific credibility to quack antivaccine views. They've clearly been co-opted by the antivaccine movement and have become true believers, so much so that Shaw is in danger of throwing away whatever scientific credibility Shaw once had. I hope they like their new friends. If they continue on this path, they'll soon be replacing their scientific colleagues for a bunch of cranks, just the way Andrew Wakefield, Mark and David Geier, and Dr. Sing Han Lee have. Soon all they'll be good for is giving aid and comfort to antivaccine cranks by churning out crap studies that can be cited over and over and over again by antivaccinationists. Oh, joy.</p>
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